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Cell functionality, driven by remarkable plasticity, is strongly influenced by mechanical forces that regulate mesenchymal stem cell (MSC) fate. This study explores the biomechanical properties of jaw periosteal cells (JPCs) and induced mesenchymal stem cells (iMSCs) under different culture conditions. We cultured both JPCs and iMSCs (n = 3) under normoxic and hypoxic environments, with and without osteogenic differentiation, and on laminin- or gelatin-coated substrates. Using atomic force microscopy, we measured cellular elasticity and Young's modulus of calcium phosphate precipitates (CaPPs) formed under osteogenic conditions. Correlation analyses between cellular stiffness, quantity of CaPP deposition, and stiffness of formed CaPPs were evaluated. The results showed that iMSCs, despite their softer cellular consistency, tended to form CaPPs of higher elastic moduli than osteogenically differentiated JPCs. Particularly under normoxic conditions, JPCs formed stronger CaPPs with lower cellular stiffness profiles. Conversely, iMSCs cultivated under hypoxic conditions on laminin-coated surfaces produced stronger CaPPs while maintaining lower cellular stiffness. We conclude that JPCs and iMSCs display distinct biomechanical responses to culture conditions. While JPCs increase cellular stiffness during osteogenic differentiation, in particular under hypoxic conditions, iMSCs exhibit a decrease in stiffness, indicating a higher resistance to lower oxygen levels. In both cell types, a lower cellular stiffness profile correlates with enhanced mineralization, indicating that this biomechanical fingerprint serves as a critical marker for osteogenic differentiation. [ABSTRACT FROM AUTHOR]

Cultivated meat refers to edible meat obtained by proliferating cells without killing livestock in a laboratory. The selection of donor animals is a crucial factor for efficient cell culture production. Hanwoo is a native Korean taurine cattle breed raised as livestock in Korea since before 2000 B.C. Cells isolated from Hanwoo, which has little genetic diversity, are expected to be advantageous in cell culture because of the existence of fewer individual differences. However, cells collected from Hanwoo are in a state where efficient culture conditions have not been established. Therefore, in this study, we investigated the effects of mimicking an in vivo environment on the proliferation and differentiation of Hanwoo muscle satellite cells. The culture conditions consisted of CON (37°C/20% O2), T1 (37°C/2% O2), T2 (39°C/20% O2), and T3 (39°C/2% O2). Cell numbers decreased and expression levels of PAX7 and MYF5 decreased at a temperature of 39°C (p < 0.05). Conversely, 2% oxygen increased the number of cells and expression levels of PAX7 and MYF5 (p < 0.05). A temperature of 39°C inhibited the proliferation of Hanwoo muscle satellite cells by reducing the expression of PAX7 and MYF5 (p < 0.05). Conversely, 2% oxygen promoted the proliferation of Hanwoo muscle satellite cells by enhancing the expression of PAX7 and MYF5 (p < 0.05). During differentiation, a temperature of 39°C improved the myotube area and fusion index (p < 0.05). The RT-qPCR and Western blotting results revealed that a culture temperature of 39°C increased expression levels of the MYH2 gene and DES and MYOG proteins (p < 0.05). Additionally, an interactive condition increased expression levels of MYOD1, DES, and MYOG genes (p < 0.05). These results indicated that a temperature of 39°C promoted the differentiation of Hanwoo muscle satellite cells by increasing DES and MYOG protein expression. Thus, the production of cultivated meat using Hanwoo muscle satellite cells is expected to be efficient under 2% oxygen for proliferation and 39°C for differentiation. [ABSTRACT FROM AUTHOR]

The bulb of Fritillaria cirrhosa D. Don is widely used for the anti-asthmatic, anti-tussive, and anti-cancer agents, etc., while the yield is limited by an endangered status, a long juvenile phase, and restricted growth habitat. Ancillary approaches to improve the bulb yield by micropropagation and bioactive metabolites production by bioreactor have not been established. Here is reported the plant regeneration, suspension cell culture, and bioactive metabolite production at different treatments. The embryogenic calli were successfully induced via the histomorphological identification. The highest proliferation times (4.11-fold) were observed with a select combination of hormones [NAA (0.2 mg/L) + 6-BA (1.0 mg/L) + GA3 (1.0 mg/L)] and culture conditions (red light and 20 °C), the highest content of imperialine (0.13 mg/g) was observed under blue light, total phenolic (0.52 mg/g) under red light, polysaccharides (36.57 mg/g) and total flavonoids (2.67 mg/g) as well as antioxidant capacity under white light. The plantlets were regenerated within 125 d from the induced embryogenic calli to acclimation and transplantation of seedlings. For the suspension cell culture, a 6.30-, 1.78-, 1.37-, and 1.51-fold increase of proliferation times, imperialine, polysaccharides, and total phenolic contents was observed at 40 d, respectively. Based on the above observations, an effective and complete in vitro approach has been proposed to regenerate plants and produce bioactive metabolites in F. cirrhosa.

Cell functionality, driven by remarkable plasticity, is strongly influenced by mechanical forces that regulate mesenchymal stem cell (MSC) fate. This study explores the biomechanical properties of jaw periosteal cells (JPCs) and induced mesenchymal stem cells (iMSCs) under different culture conditions. We cultured both JPCs and iMSCs (n = 3) under normoxic and hypoxic environments, with and without osteogenic differentiation, and on laminin- or gelatin-coated substrates. Using atomic force microscopy, we measured cellular elasticity and Young’s modulus of calcium phosphate precipitates (CaPPs) formed under osteogenic conditions. Correlation analyses between cellular stiffness, quantity of CaPP deposition, and stiffness of formed CaPPs were evaluated. The results showed that iMSCs, despite their softer cellular consistency, tended to form CaPPs of higher elastic moduli than osteogenically differentiated JPCs. Particularly under normoxic conditions, JPCs formed stronger CaPPs with lower cellular stiffness profiles. Conversely, iMSCs cultivated under hypoxic conditions on laminin-coated surfaces produced stronger CaPPs while maintaining lower cellular stiffness. We conclude that JPCs and iMSCs display distinct biomechanical responses to culture conditions. While JPCs increase cellular stiffness during osteogenic differentiation, in particular under hypoxic conditions, iMSCs exhibit a decrease in stiffness, indicating a higher resistance to lower oxygen levels. In both cell types, a lower cellular stiffness profile correlates with enhanced mineralization, indicating that this biomechanical fingerprint serves as a critical marker for osteogenic differentiation.

The environmental conditions were optimized to produce the enniatin H, I, and MK1688 by Fusarium strain on cereal grain exhibiting anti-carcinogenic potential against MES-SA (human uterine sarcoma cell line), HCT15 (human colorectal carcinoma cancer cell line), and their multidrug resistance sublines. From the statistical optimization by response surface methodology, the optimal condition of independent variables affecting the response variables were 20.85 °C (temperature), 46.85% (w/w, initial moisture content), and 18.42 days (growth time) for ENN H; 23.31 °C, 44.15% (w/w) and 17.23 days for ENN I; 23.08 °C, 43.97% (w/w) and 17.06 days for ENN MK1688. In case of cytotoxic effects, ENNs significantly suppressed growth of cancer cell lines without multidrug resistance, and ENN I inhibited growth of cancer cell lines most strongly. These data will provide valuable point to produce the cyclic hexadepsipeptide exhibiting anti-carcinogenic potential from Fusarium strains. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to explore the optimal medium composition and culture conditions for obtaining the maximum viable count of Brevibacillus laterosporus. Methods: Based on establishing the correlation regression equation between the MTT colorimetric method and plate counting method, the optimal medium composition (carbon source, nitrogen source, inorganic salt) and fermentation conditions (initial pH, temperature, inoculation amount, KH2PO4) for obtaining the maximum viable count of B. laterosporus were optimized. Results: A significant linear correlation for viable count determination between MTT colorimetry and plate counting (R2>0.999) was found; Maltose, CaCl2, initial pH and KH2PO4 were the significant factors. The optimal fermentation condition was 8.75 g/L maltose, 0.17 g/L CaCl2, 7.07 initial pH and 3.73 g/L KH2PO4. Under the control of these conditions, the number of viable counts was 8.12×108 CFU/mL. However, no significant difference from the theoretical number (8.25×108 CFU/mL) was found. Conclusion: Based on MTT colorimetry and response surface methodology, the culture conditions for the maximum viable count of B. laterosporus were optimized and the number of viable counts was higher than before optimization increased by 3.02 times.

In the last four decades, the assisted reproductive technology (ART) field has witnessed advances, resulting in improving pregnancy rates and diminishing complications, in particular reduced incidence of multiple births. These improvements are secondary to advanced knowledge on embryonic physiology and metabolism, resulting in the ability to design new and improved culture conditions. Indeed, the incubator represents only a surrogate of the oviduct and uterus, and the culture conditions are only imitating the physiological environment of the female reproductive tract. In vivo, the embryo travels through a dynamic and changing environment from the oviduct to the uterus, while in vitro, the embryo is cultured in a static fashion. Importantly, while culture media play a critical role in optimising embryo development, a large host of additional factors are equally important. Additional potential variables, including but not limited to pH, temperature, osmolality, gas concentrations and light exposure need to be carefully controlled to prevent stress and permit optimal implantation potential. This manuscript will provide an overview of how different current culture conditions may affect oocyte and embryo viability with particular focus on human literature. [ABSTRACT FROM AUTHOR]

Third-generation biomass production utilizing microalgae exhibits sustainable and environmentally friendly attributes, along with significant potential as a source of physiologically active compounds. However, the process of screening and localizing strains that are capable of producing high-value-added substances necessitates a significant amount of effort. In the present study, we have successfully isolated the indigenous marine diatom Odontella aurita OAOSH22 from the east coast of Korea. Afterwards, comprehensive analysis was conducted on its morphological, molecular, and biochemical characteristics. In addition, a series of experiments was conducted to analyze the effects of various environmental factors that should be considered during cultivation, such as water temperature, salinity, irradiance, and nutrients (particularly nitrate, silicate, phosphate, and iron). The morphological characteristics of the isolate were observed using optical and electron microscopes, and it exhibited features typical of O. aurita. Additionally, the molecular phylogenetic inference derived from the sequence of the small-subunit 18S rDNA confirmed the classification of the microalgal strain as O. aurita. This isolate has been confirmed to contain 7.1 mg g−1 dry cell weight (DCW) of fucoxanthin, a powerful antioxidant substance. In addition, this isolate contains 11.1 mg g−1 DCW of eicosapentaenoic acid (EPA), which is one of the nutritionally essential polyunsaturated fatty acids. Therefore, this indigenous isolate exhibits significant potential as a valuable source of bioactive substances for various bio-industrial applications. [ABSTRACT FROM AUTHOR]

We aimed to determine the optimal conditions for the mass culture of rotifers, which can be used as feed for cold-water fish species at low temperatures. The growth and specific growth rates (SGRs) of rotifers were assessed considering water temperature, salinity, density, dissolved oxygen (DO) levels, and the amount of Chlorella supplied as feed. The growth of rotifers was higher at 15 °C than at 10 °C and at salinities of ~11–17 psu. Initial inoculation densities of 500 and 700 individuals/mL resulted in the highest rotifer density, and SGR was highest at 100 individuals/mL. DO concentration did not significantly affect the growth and SGRs of rotifers. Enrichment with fatty acids is important to supplement the diet of cold-water fish species. Highly unsaturated fatty acid content increased with enrichment time to 14.04 ± 0.86% at 12 h and 15.58 ± 2.20% at 24 h. Thus, the optimal conditions for rotifer mass culture are a water temperature of 15 °C, salinity of 11–17 psu, initial inoculation density of 300–500 individuals/mL, DO concentration of 8 mg/L or more, and Chlorella supply at 7.5 × 1012 cells/mL. Therefore, the present study suggests optimal culture conditions of rotifers at low temperatures for breeding cold-water fish species. [ABSTRACT FROM AUTHOR]

The growth and gas production test for Escherichia coli in the microbiological examination of food additives is stipulated in the ninth edition of Japan's Specifications and Standards for Food Additives (JSFA) and described as a part of the "Confirmation Test for Escherichia coli" in "Microbial Limit Tests" in the same manuscript. The growth and gas production test for E. coli indicated that the positive or negative of "gas production and/or turbidity" in EC broth should be confirmed after incubating at 45.5±0.2°C for 24±2 h. If both gas production and turbidity are negative, the culture is additionally incubated up to 48±2 h to determine E. coli contamination. The internationally referenced Bacteriological Analytical Manual of the U.S. FDA had revised the incubation temperature in tests for coliforms and E. coli from 45.5±0.2°C to 44.5±0.2°C in 2017. Therefore, we conducted research in anticipation of this temperature change being reflected in the microbiological examination of the JSFA. We used seven EC broth products and six food additives across eight products that are available in Japan in order to compare the growth and gas production at temperatures of 45.5±0.2°C and 44.5±0.2°C of E. coli NBRC 3972, which is designated as the test strain in JSFA. Both with/without food additives, the number of EC broth products in which medium turbidity and gas production by the strain were positive in three out of three tubes at all test times was greater at 44.5±0.2°C than at 45.5±0.2°C. These results suggest that the growth and gas production test for E. coli could be more appropriately conducted by incubation at 44.5±0.2°C in the "Confirmation Test for Escherichia coli" for E. coli in the JSFA in comparison to 45.5±0.2°C. Furthermore, there were differences in the growth and gas production of E. coli NBRC 3972 depending on the EC broth product used. Therefore, the importance of "Media growth promotion test" and "Method suitability test" in the ninth edition of the JSFA should be emphasized. [ABSTRACT FROM AUTHOR]

Microbial oil has been gaining considerable attention from researchers recently as renewable and ecofriendly oil and its potential as feedstock for food industry and biodiesel industry. In this context, we have earlier demonstrated production of microbial oil and exopolysaccharide (EPS) from the yeast Sporidiobolus pararoseus JD-2. In this study, we explored increasing its production by optimizing the culture condition and nutrition. As expected, culture temperature and dissolved oxygen (DO) are the contributing factors for co-producing microbial oil and EPS, in which 28℃ and lower quantum (i.e., 30 mL/500 mL) show the best conditions in shake-flasks fermentation. By contrast, the initial pH from 4 to 8 has no obvious effect on producing microbial oil and EPS. In addition, the culture nutrition (i.e., carbon/nitrogen source) were also discussed, and indicating that 20 g/L of corn steep liquor and 60 g/L of glucose are beneficial to produce microbial oil and EPS (i.e., 34.1±1.2 g/L and 11.5±0.2 g/L, respectively). Meanwhile, the residue glucose should be maintained at 20 g/L, in which the highest production of microbial oil and EPS was obtained (i.e., 34.6±1.7 g/L and 11.7±0.8 g/L, respectively). The biomass, microbial oil and EPS were further increased during optimizing the DO level, which reached to 67.8±2.1 g/L, 34.7±0.6 g/L and 11.8±0.5 g/L during maintaining DO level at 20-30%, respectively. The results suggest that appropriate culture condition and nutrition considerably improve the fermentation performance of S. pararoseus JD-2 and significantly increase co-production of microbial oil and EPS (by 11.2 and 8.3%, respectively) compared to the un-optimized fermentation. [ABSTRACT FROM AUTHOR]

Recombinant mammalian host cell lines, in particular CHO and HEK293 cells, are used for the industrial production of therapeutic proteins. Despite their well-known genomic instability, the control mechanisms that enable cells to respond to changes in the environmental conditions are not yet fully understood, nor do we have a good understanding of the factors that lead to phenotypic shifts in long-term cultures. A contributing factor could be inherent diversity in transcriptomes within a population. In this study, we used a full-length coverage single-cell RNA sequencing (scRNA-seq) approach to investigate and compare cell-to-cell variability and the impact of standardized and homogenous culture conditions on the diversity of individual cell transcriptomes, comparing suspension CHO-K1 and adherent HEK293FT cells. Our data showed a critical batch effect from the sequencing of four 96-well plates of CHO-K1 single cells stored for different periods of time, which was and may be therefore identified as a technical variable to consider in experimental planning. Besides, in an artificial and controlled culture environment such as used in routine cell culture technology, the gene expression pattern of a given population does not reveal any marker gene capable to disclose relevant cell population substructures, both for CHO-K1 cells and for HEK293FT cells. The variation observed is primarily driven by the cell cycle. [Display omitted] • scRNA-seq of CHO-K1 suspension and HEK293FT adherent cultures shows cellular homogeneity and lack of clear subpopulations. • The minor source of heterogeneity of HEK293FT attributable to the cell cycle may be due to the limited cells examined. • 53 highly variable genes shared between cell lines display the transcriptomes as close neighbours in the 2D-embedding plot. • The holding time of printed plates may affect the quality of the sequencing data and the variance in the data sets. [ABSTRACT FROM AUTHOR]

Alzheimer's disease (AD) is a common age-related neurodegenerative disease characterized by progressive cognitive decline and irreversible memory impairment. Currently, several studies have failed to fully elucidate AD's cellular and molecular mechanisms. For this purpose, research on related cellular models may propose potential predictive models for the drug development of AD. Therefore, many cells characterized by neuronal properties are widely used to mimic the pathological process of AD, such as PC12, SH-SY5Y, and N2a, especially the PC12 pheochromocytoma cell line. Thus, this review covers the most systematic essay that used PC12 cells to study AD. We depict the cellular source, culture condition, differentiation methods, transfection methods, drugs inducing AD, general approaches (evaluation methods and metrics), and in vitro cellular models used in parallel with PC12 cells. [ABSTRACT FROM AUTHOR]

Chronic inflammation caused by aging, obesity, and lifestyle disturbances can lead to the production of inflammatory cytokines and insulin resistance, reducing glucose and lipid metabolism. Lactic acid bacteria (LAB) have various bioactivities, and certain types of LAB have been reported to exhibit anti-inflammatory effects. We hypothesized that LAB strains, which can strongly induce the production of anti-inflammatory cytokines by immune cells in the intestinal tract, may improve glucose and lipid metabolism by suppressing chronic inflammation. We selected Lactiplantibacillus plantarum OLL2712 (OLL2712) from the LAB library owned by Meiji Co., Ltd. based on its ability to induce the production of interleukin-10 (IL-10), optimized the culture conditions of OLL2712 for industrial applications, and verified the efficacy of the strain in animal and clinical studies. The results showed that OLL2712 bacterial cells in the exponential phase had notably higher anti-inflammatory properties than the cells in the stationary phase and led to the inhibition of chronic inflammation and improvement of glucose and lipid metabolism in animal studies. Two randomized controlled trials consisting of healthy adults with elevated blood glucose levels or body mass indices (BMIs) also showed that the intake of OLL2712 suppressed the aggravation of chronic inflammation and improved glucose and lipid metabolism. This review identified a novel LAB strain that may contribute to diabetes and obesity prevention and demonstrated its clinical efficacy. In addition, the mechanism of action of this LAB strain through the intestinal immune system was partially elucidated, and the importance of optimizing the culture conditions of LAB was clarified. [ABSTRACT FROM AUTHOR]

Third-generation biomass production utilizing microalgae exhibits sustainable and environmentally friendly attributes, along with significant potential as a source of physiologically active compounds. However, the process of screening and localizing strains that are capable of producing high-value-added substances necessitates a significant amount of effort. In the present study, we have successfully isolated the indigenous marine diatom Odontella aurita OAOSH22 from the east coast of Korea. Afterwards, comprehensive analysis was conducted on its morphological, molecular, and biochemical characteristics. In addition, a series of experiments was conducted to analyze the effects of various environmental factors that should be considered during cultivation, such as water temperature, salinity, irradiance, and nutrients (particularly nitrate, silicate, phosphate, and iron). The morphological characteristics of the isolate were observed using optical and electron microscopes, and it exhibited features typical of O. aurita. Additionally, the molecular phylogenetic inference derived from the sequence of the small-subunit 18S rDNA confirmed the classification of the microalgal strain as O. aurita. This isolate has been confirmed to contain 7.1 mg g−1 dry cell weight (DCW) of fucoxanthin, a powerful antioxidant substance. In addition, this isolate contains 11.1 mg g−1 DCW of eicosapentaenoic acid (EPA), which is one of the nutritionally essential polyunsaturated fatty acids. Therefore, this indigenous isolate exhibits significant potential as a valuable source of bioactive substances for various bio-industrial applications.

乙酰辅酶A羧化酶是谷氨酸棒杆菌中调控脂肪酸合成的关键酶。为了研究乙酰辅酶A羧化酶α亚基（accBC）基因对脂肪酸合成的影响，以accBC表达的菌株重组谷氨酸棒杆菌G-BC为材料，研究了诱导剂异丙基-β-D-硫代半乳糖苷（IPTG）浓度、培养时间、培养温度对重组谷氨酸棒杆菌G-BC脂肪酸产量的影响，并通过响应面法优化了重组谷氨酸棒杆菌G-BC产脂肪酸的培养条件，同时与原始菌株脂肪酸产量进行了对比。结果表明：重组谷氨酸棒杆菌G-BC产脂肪酸的最佳条件为诱导剂浓度1 mmol/L 、培养时间20 h、培养温度32 ℃，在此条件下脂肪酸产量为34.56 mg/g（以菌体干质量计），比原始菌株提高了1.68倍。说明乙酰辅酶A羧化酶α亚基对于谷氨酸棒杆菌合成脂肪酸有促进作用。 Acetyl-CoA carboxylase is a key enzyme regulating fatty acid synthesis in Corynebacterium glutamicum. In order to study the effect of acetyl-CoA carboxylase α subunit (accBC) gene on fatty acid synthesis,the recombinant Corynebacterium glutamicum G-BC(expression strain of accBC) was used as raw material, the effects of inducer(isopropyl-β-D-thiogalactopyranoside,IPTG) concentration, culture time and culture temperature on the yield of fatty acid were studied. Moreover, the culture conditions for fatty acid production of recombinant Corynebacterium glutamicum G-BC were optimized by response surface methodology. The fatty acid yield of the original strain and the recombinant strain was also compared.The results showed that the optimal culture conditions were obtained as follows: inducer concentration 1 mmol/L, culture time 20 h, and culture temperature 32 ℃. Under these conditions, the yield of fatty acid was 34.56 mg/g（based on dry weight of bacteria）, which was 1.68 times higher than original strain. The accBC can promote the synthesis of fatty acid by Corynebacterium glutamicum.

The US Environmental Protection Agency listed phenolic compounds as priority pollutants, and their occurrence in water systems poses serious health risks to humans and other living species. The culture conditions are extremely crucial for microbial growth, enzymatic and cellular metabolic activities. Thus, the effects of different culture parameters like pH, temperature (°C), and agitation speed (RPM) on the indigenous Rhodococcus pyridinivorans strain PDB9T NS-1 were modeled using response surface methodology (RSM) and central composite design (CCD). The results showed that the main effect of pH and interaction between pH and agitation speed had a significant to a moderately significant effect on phenol degradation by the indigenous R. pyridinivorans strain PDB9T NS-1. Almost complete phenol degradation was obtained at an optimal setting of pH 7.5, 187 rpm, and 34 °C in 18 h. Growth and phenol degradation kinetics of the actinomycetes was examined in a batch shake flask under the optimized conditions. The growth of Rhodococcus species at varying initial levels of the phenol followed a Pamukoglu and Kargi substrate inhibition model with half-saturation constant (Ks) of 54.21 mg/l, maximum specific growth rate (µmax) of 0.169 (1/h), substrate inhibition constant (Ksi) of 243.26 mg/l. The results obtained from the optimization of culture conditions and growth kinetics by the indigenous R. Pyridinivorans strain PDB9T NS-1 suggest the potential of the system in the treatment of phenolic wastewater. [ABSTRACT FROM AUTHOR]

Background/Aim: Chimeric antigen receptor (CAR) T cell therapy targeting CD20 has the potential to become a promising novel treatment for canine B cell lymphoid malignancy. However, the optimal approach for producing potent CAR-T cells with favorable phenotype for dogs remains unknown. In this study, we assessed several culture conditions and their effects on the phenotypic characteristics of CD20-CAR-T cells. Materials and Methods: Canine CAR-T cells were generated by incubating with several mitogens in the presence or absence of Akt inhibitor. Gene transduction efficiency and phenotypic characteristics were determined by flow cytometry. Results: Comparison of several kinds of mitogens revealed that stimulation with phytohemagglutinin has high transduction efficacy, whereas stimulation with concanavalin A was superior in memory T cell formation. Akt inhibition at the initial stage of CAR-T production tended to enhance transduction efficiency and memory T cell formation. Conclusion: This study provides a significant insight into the understanding of the ex vivo expansion of canine T cells in adoptive immunotherapy. [ABSTRACT FROM AUTHOR]

Fucoxanthin is one of the light-harvesting pigments in brown microalgae, which is increasingly gaining attention due to its numerous health-promoting properties. Currently, the production of microalgal fucoxanthin is not yet feasible from an economic perspective. However, the cultivation of microalgae at favourable conditions holds great potential to increase the viability of this fucoxanthin source. Hence, this study aimed to review the fucoxanthin production of microalgae under different conditions systematically. A literature search was performed using the Web of Science, Scopus and PubMed databases. A total of 188 articles were downloaded and 28 articles were selected for the current review by two independent authors. Microalgae appeared to be a more reliable fucoxanthin source compared to macroalgae. Overall, a consensus fucoxanthin production condition was obtained and proposed: light intensity ranging from 10 to 100 µmol/m2/s could achieve a higher fucoxanthin content. However, the optimal light condition in producing fucoxanthin is species-specific. The current review serves as an antecedent by offering insights into the fucoxanthin-producing microalgae response to different culture factors via a systematic analysis. With the current findings and recommendations, the feasibility of producing fucoxanthin commercially could be enhanced and possibly achieve practical and sustainable fucoxanthin production.

The species Pantoea agglomerans includes strains that are agronomically relevant for their growth-promoting or biocontrol traits. Molecular analysis demonstrated that the IPDC pathway involved in the conversion of tryptophan (Trp) to indole-3-acetic acid (IAA) is highly conserved among P. agglomerans strains at both gene and protein levels. Results also indicated that the promoter region controlling the inducible expression of ipdC gene differs from the model system Enterobacter cloacae, which is in accordance with the observation that P. agglomerans accumulates higher levels of IAA when cells are collected in the exponential phase of growth. To assess the potential applications of these microorganisms for IAA production, P. agglomerans C1, an efficient auxin-producer strain, was cultivated in 5 L fermenter so as to evaluate the effect of the medium formulation, the physiological state of the cells, and the induction timing on the volumetric productivity. Results demonstrated that higher IAA levels were obtained by using a saline medium amended with yeast extract and saccharose and by providing Trp, which acts both as a precursor and an inducer, to a culture in the exponential phase of growth. Untargeted metabolomic analysis revealed a significant effect of the carbon source on the exometabolome profile relative to IAA-related compounds and other plant bioactive signaling molecules. The IAA-enriched metabolites secreted in the culture medium by P. agglomerans C1 were used as plant biostimulants to run a series of trials at a large-scale nursery farm. Tests were carried out with in vitro and ex vitro systems following the regular protocols used for large-scale plant tree agamic propagation. Results obtained with 4,540 microcuttings of Prunus rootstock GF/677 and 1,080 plantlets of Corylus avellana L. showed that metabolites from strain C1 improved percentage of rooted-explant, number of adventitious root formation, plant survival, and quality of plant as vigor, with an increase in the leaf area between 17.5 and 42.7% compared to IBA-K (indole-3-butyric acid potassium salt)–treated plants.

Abstract Utilization of algicidal bacteria as a biological agent have been receiving significant interest for controlling harmful algal blooms. While various algicidal bacterial strains have been identified, limited studies have explored the influence of bacterial culture conditions on its algicidal activity. Here, the effect of oxygen on the algicidal activity of a novel bacterium JK12, against a model diatom, Phaeodactylum tricornutum (P. tricornutum) was studied. Strain JK12 showed high algicidal activity against P. tricornutum and was identified as Pseudomonas chlororaphis (P. chlororaphis) by 16S ribosomal RNA gene analysis. JK12 culture supernatant exhibited strong algicidal activity while washed JK12 cells showed no obvious activity, indicating that JK12 indirectly attacks algae by secreting extracellular algicidal metabolites. Micro-aerobic culture condition dramatically enhanced the algicidal activity of JK12 by 50%, compared to that cultured under aerobic condition in 24 h. Extracellular metabolomic profiling of JK12 using gas chromatography–mass spectrometry and liquid chromatography–mass spectrometry analysis revealed significantly higher amounts of allantoic acid, urocanic acid, cytidine 2′,3′-cyclic phosphate, uridine 2′,3′-cyclic phosphate, and chlorinated tryptophan in the micro-aerobic culture. This is the first report to demonstrate the important role of oxygen on the algicidal activity of a non-pathogenic strain P. chlororaphis. In addition, the metabolomics analysis provided insights into the algicidal mechanism of P. chlororaphis.

Abstract Background Aberrant DNA methylation patterns of genes required for development are common in in vitro produced embryos. In this regard, we previously identified altered DNA methylation patterns of in vivo developed blastocysts from embryos which spent different stages of development in vitro, indicating carryover effects of suboptimal culture conditions on epigenetic signatures of preimplantation embryos. However, epigenetic responses of in vivo originated embryos to suboptimal culture conditions are not fully understood. Therefore, here we investigated DNA methylation patterns of in vivo derived bovine embryos subjected to in vitro culture condition before, during or after major embryonic genome activation (EGA). For this, in vivo produced 2-, 8- and 16-cell stage embryos were cultured in vitro until the blastocyst stage and blastocysts were used for genome-wide DNA methylation analysis. Results The 2- and 8-cell flushed embryo groups showed lower blastocyst rates compared to the 16-cell flush group. This was further accompanied by increased numbers of differentially methylated genomic regions (DMRs) in blastocysts of the 2- and 8-cell flush groups compared to the complete in vivo control ones. Moreover, 1623 genomic loci including imprinted genes were hypermethylated in blastocyst of 2-, 8- and 16-cell flushed groups, indicating the presence of genomic regions which are sensitive to the in vitro culture at any stage of embryonic development. Furthermore, hypermethylated genomic loci outnumbered hypomethylated ones in blastocysts of 2- and 16-cell flushed embryo groups, but the opposite occurred in the 8-cell group. Moreover, DMRs which were unique to blastocysts of the 2-cell flushed group and inversely correlated with corresponding mRNA expression levels were involved in plasma membrane lactate transport, amino acid transport and phosphorus metabolic processes, whereas DMRs which were specific to the 8-cell group and inversely correlated with corresponding mRNA expression levels were involved in several biological processes including regulation of fatty acids and steroid biosynthesis processes. Conclusion In vivo embryos subjected to in vitro culture before and during major embryonic genome activation (EGA) are prone to changes in DNA methylation marks and exposure of in vivo embryos to in vitro culture during the time of EGA increased hypomethylated genomic loci in blastocysts.

Drynaria bonii H. Christ is a medicinal plant that has traditionally been used for the treatment of chronic diseases and tuberculosis. However, D. bonii has not yet been cultivated or propagated. In this study, we optimized the conditions for the spore culture of D. bonii to produce sporophytes. First, we evaluated the effects of various factors, such as temperature, light intensity, potting material, and pH, on spore germination and early gametophyte development. Under optimal culture conditions, 95–99% of the spores germinated within 2 weeks of culture. Next, based on these results, we established a culture system for gametophyte development and sporophyte production. Mature gametophytes first appeared 5 months post-germination (mpg), and at 12 mpg, the rate of production of heart-shaped gametophytes reached 56.6%. Sporophytes at the early first-leaf stage first appeared at 6 mpg, and at 12 mpg, the rate of formation of sporophytes reached 15.4%. Interestingly, 6.3% of all gametophytes produced multiple sporophytes. Additionally, 42–62% of the gametophytes also produced multiple secondary gametophytes, indicating a high potential of D. bonii gametophytes to generate new gametophytes and, subsequently, sporophytes. When transferred to new trays or pots, sporophytes grew well and showed 100% survival. Overall, we conclude that this spore culture system can be successfully used for the propagation of D. bonii sporophytes. [ABSTRACT FROM AUTHOR]

The species Pantoea agglomerans includes strains that are agronomically relevant for their growth-promoting or biocontrol traits. Molecular analysis demonstrated that the IPDC pathway involved in the conversion of tryptophan (Trp) to indole-3-acetic acid (IAA) is highly conserved among P. agglomerans strains at both gene and protein levels. Results also indicated that the promoter region controlling the inducible expression of ipdC gene differs from the model system Enterobacter cloacae , which is in accordance with the observation that P. agglomerans accumulates higher levels of IAA when cells are collected in the exponential phase of growth. To assess the potential applications of these microorganisms for IAA production, P. agglomerans C1, an efficient auxin-producer strain, was cultivated in 5 L fermenter so as to evaluate the effect of the medium formulation, the physiological state of the cells, and the induction timing on the volumetric productivity. Results demonstrated that higher IAA levels were obtained by using a saline medium amended with yeast extract and saccharose and by providing Trp, which acts both as a precursor and an inducer, to a culture in the exponential phase of growth. Untargeted metabolomic analysis revealed a significant effect of the carbon source on the exometabolome profile relative to IAA-related compounds and other plant bioactive signaling molecules. The IAA-enriched metabolites secreted in the culture medium by P. agglomerans C1 were used as plant biostimulants to run a series of trials at a large-scale nursery farm. Tests were carried out with in vitro and ex vitro systems following the regular protocols used for large-scale plant tree agamic propagation. Results obtained with 4,540 microcuttings of Prunu s rootstock GF/677 and 1,080 plantlets of Corylus avellana L. showed that metabolites from strain C1 improved percentage of rooted-explant, number of adventitious root formation, plant survival, and quality of plant as vigor, with an increase in the leaf area between 17.5 and 42.7% compared to IBA-K (indole-3-butyric acid potassium salt)–treated plants. [ABSTRACT FROM AUTHOR]

This study aimed to examine the dyes production of mixed Aspergillus and Paecilomyces on different culture conditions and evaluate the combination between culture and dyeing conditions on cloth color by the fungal dyes. The different media consist of mineral salt glucose (MSG) medium and mineral salt sucrose (MSS) medium were evaluated. Additionally, different culture conditions such as C and N sources, incubation temperature, and pH of the media were evaluated together with variation of dyeing conditions including pH and mordants. The result showed that different culture conditions on media and the combination of those with dyeing conditions produce distinction colors on the fabric. The variation of dyed colors on the fabric was related to the dyes concentration produced by combination treatment of culture condition and dyeing pH. However, the effect was not observed on the combination between culture condition and mordants. Combination of culture and dyeing conditions on cloth color dyed with mixed fungi dyes generated various cloth colors, adding a color variation on textile dyeing. [ABSTRACT FROM AUTHOR]

ABSTRACT Many microalgae are rich in lipid. Due to their low growth rates in the autotrophic culture, the best alternative is to cultivate cells under different conditions such as heterotrophic or mixotrophic, which results in the highest yield of biomass and lipid in the shortest duration. In this study, Dunaliela bardawil (UTEX 2538) green microalgae was cultivated under different culture conditions, autotrophic, heterotrophic and mixotrophic, and effects on cell concentration, lipid production and reactive oxygen species (ROS), total chlorophyll and beta-carotene concentrations were investigated. Due to very low cellular growth, this alga is not recommended for heterotrophic culture. In terms of mixotrophic conditions containing different concentrations of glucose or acetate, the highest cell growth and lipid production in 60mM glucose was similar to the control (autotrophic condition), while the concentrations of chlorophyll and beta-carotene decreased. However, at all concentrations of acetate, a slight increase in cell growth was achieved, while the lipid content decreased. Additionally, the concentrations of chlorophyll and beta-carotene increased. A positive correlation was observed between beta-carotene biosynthesis and lipid production, as well as levels of reactive species of oxygen and lipid production in the presence of glucose and acetate. This study showed that for D. bardawil the mixotrophic culture with 60mM glucose was the most suitable type of culture for increasing lipid content and cell growth rates in less time (one week).

The objective of this study was the development of bioprocess for enhanced biomass production of probiotic Bifidobacterium bifidum. In the first process optimization step in Erlenmeyer flasks cultures, different experiments were conducted to study the effect of inoculum volume, inoculum age, temperature and pH of the growth medium on the kinetics of cell growth. In Erlenmeyer flasks cultures, the maximal biomass production was observed with 1% inoculum of 6 log hours at 37°C, and optimal pH of initial media was found to be 6.0. Further positive development in biomass production was observed by scaling up the fermentation process to stirred tank bioreactor. Fermentation was carried out in 2L stirred tank bioreactor, with agitation of 100 rpm and constant temperature of 37°C. The batch culture produced higher biomass of 34.1 g wet cell weight g/l in 12 log hours and viable counts (2.5 × 109 CFU/ml) compared to Erlenmeyer flasks. In conclusion, batch cultivation in the 2 l bioreactor with this growth medium under optimal conditions gives enhanced biomass production. However, based on our end result, high-cell density fed-batch and pH control strategies are recommended for the commercial production of B. bifidum as a probiotic. [ABSTRACT FROM AUTHOR]

Combination of microalgae cultivation and piggery wastewater treatment has become a hot topic in recent years. Nutrient removal from aerated piggery wastewater (APW) by Desmodesmus sp. CHX1 and the optimization of cultivation conditions were investigated in this study. Results indicated that Desmodesmus sp. CHX1 showed an efficient growth in APW, with specific growth rate of 0.26–0.56 d−1. The biomass yield based on nutrient consumption was 9.65 g biomass/g NH4–N and 209.15 g biomass/g total phosphorus (TP) respectively. Desmodesmus sp. CHX1 performed well in nutrient removal from APW, with ammonium nitrogen (NH4–N) and TP removal efficiency (RE) of 78.46% and 91.66% respectively after 7 days of culture. Nutrient removal process fitted the pseudo-first-order kinetic equations well, with removal rate (RR) constant of 0.24 d−1 for NH4–N and 0.28 d−1 for TP. The optimum conditions for nutrient removal from APW by Desmodesmus sp. CHX1 were light intensity of 150 μmol photons m−2 s−1 in the photoperiod for 24 h when the temperature was set at 35°C with alga cell inoculation concentration of 30%. The removal efficiencies of NH4–N and TP were 88.26% and 95.06% respectively under the optimal conditions after 7 days of culture. Our results can be a good reference for enhancement of microalga production and the nutrient RE and further extend the application of the large-scale piggery wastewater treatment under a controlled environment. [ABSTRACT FROM AUTHOR]

Aims: The aims of this study were to identify the structure of antibacterial lipopeptide (LP) produced by Bacillus amyloliquefaciens strain FJAT‐2349, to analyse the effects of the culture medium and temperature on LP production and to assess the biocontrol efficiency of the LPs against tomato bacterial wilt. Methods and Results: Lipopeptides were extracted by acid precipitation and resolved in methanol and their structure was identified through LC‐QTOF‐MS/MS method. The antibacterial activities of the LPs were evaluated through inhibition zone experiments. The biocontrol efficiency of the LPs against tomato bacterial wilt was examined by a pot test. The LPs were composed of iturin (C14–C17 iturin A), fengycin (C14/C16 fengycin A, C14 fengycin B2, C16 fengycin A2/B2, C16–C17 fengycin B, C15 fengycin A derivatives and C15 fengycin B derivatives) and surfactin (C12–C16 surfactin A). Moreover, the composition of the LPs was significantly influenced by the culture medium and temperature; the contents of iturin, fengycin and surfactin varied within the range from 0·41–5·89, 4·54–181·67 and 2·05–19·65 mg l−1 in the different culture media and from 0·39–11·04, 1·45–215·14 and 7·79–24·43 mg l−1 under different culture temperatures respectively. The results indicated that the contents of the LP mixture, fengycin and surfactin secreted from FJAT‐2349 all decreased along with an increasing culture temperature. The fermentation supernatants and LP extracts had the strongest inhibition activities of Ralstonia solanacearum when strain FJAT‐2349 was incubated at 25°C using potato dextrose broth as the culture medium among all the assayed culture conditions. The purified fengycin was found to be the active antibacterial compound against R. solanacearum, but the purified surfactin was not. The pot experiments demonstrated that the LPs secreted from the strain FJAT‐2349 could effectively control the tomato bacterial wilt with a biocontrol efficiency of 97·6%. Conclusions: The LPs secreted from strain FJAT‐2349 could serve as potential biocontrol agents against tomato bacterial wilt. Significance and Impact of the Study: The LPs exhibited good potential applications in the biocontrol of tomato bacterial wilt. [ABSTRACT FROM AUTHOR]

Crop culture conditions are one of the important interfaces between food, the environment, and health, and an essential research area for maintaining social-ecological integrity. In recent years, it has been reported that the difference in culture conditions between monoculture with external inputs (in cultura) and self-organized ecological niches (in natura) is significant for the resulting physiological property of plants. It has also been suggested that there exist metabolic proxies in various foods that can separate these two culture conditions, which does not depend on a single component but on the distribution of various compounds. However, little has been studied in a time series of replicated production to quantify the reproducibility of these metabolomic features associated with culture conditions. In this study, we obtained metabolome data of coarse green tea (Camellia sinensis) grown in the same region in Japan under both in cultura and in natura culture conditions over the course of six years, and constructed a list of multiple components that separated the effects of culture conditions by statistical analysis, and estimated the metabolic functions of the compounds that contributed to the separation. The results suggest that naturally grown samples are rich in allelochemicals, such as phytochemicals, alkaloids, phenylpropanoids, steroids, as well as the compounds related to microorganisms and vitamin B6 that imply the interactions with the soil microbiome. The estimated physiological functions of the distinctive compounds suggest that the in natura crop production is not only beneficial with known properties of maintaining ecosystem health such as soil functions and pathogen control, but also for the augmentation of the plant secondary metabolites that support long-term health protective effects.

Upon activation T cells engage glucose metabolism to fuel the costly effector functions needed for a robust immune response. Consequently, the availability of glucose can impact on T cell function. The glucose concentrations used in conventional culture media and common metabolic assays are often artificially high, representing hyperglycaemic levels rarely present in vivo. We show here that reducing glucose concentration to physiological levels in culture differentially impacted on virus-specific compared to generically activated human CD8 T cell responses. In virus-specific T cells, limiting glucose availability significantly reduced the frequency of effector-cytokine producing T cells, but promoted the upregulation of CD69 and CD103 associated with an increased capacity for tissue retention. In contrast the functionality of generically activated T cells was largely unaffected and these showed reduced differentiation towards a residency phenotype. Furthermore, T cells being cultured at physiological glucose concentrations were more susceptible to viral infection. This setting resulted in significantly improved lentiviral transduction rates of primary cells. Our data suggest that CD8 T cells are exquisitely adapted to their niche and provide a reminder of the need to better mimic physiological conditions to study the complex nature of the human CD8 T cell immune response.

Much interest has been directed towards stem cells, both in basic and translational research, to understand basic stem cell biology and to develop new therapies for many disorders. In general, stem cells can be cultured with relative ease, however, most common culture methods for stem cells employ 2D techniques using plastic. These cultures do not well represent the stem cell niches in the body, which are delicate microenvironments composed of not only stem cells, but also supporting stromal cells, extracellular matrix, and growth factors. Therefore, researchers and clinicians have been seeking optimal stem cell preparations for basic research and clinical applications, and these might be attainable through 3D culture of stem cells. The 3D cultures recapitulate the in vivo cell-to-cell and cell-to-matrix interactions more effectively, and the cells in 3D cultures exhibit many unique and desirable characteristics. The culture of stem cells in 3D may employ various matrices or scaffolds, in addition to the cells, to support the complex structures. The goal of this Special Issue is to bring together recent research on 3D cultures of various stem cells to increase the basic understanding of stem cells and culture techniques, and also highlight stem cell preparations for possible novel therapeutic applications.

Three polysaccharides (WL-CPS-1, NaCl-CPS-1 and Glu-CPS-1) were extracted and purified from Nostoc flagelliforme under normal, salt stress and mixotrophic culture conditions respectively. Their physicochemical properties and antioxidant activities were investigated. WL-CPS-1, NaCl-CPS-1 and Glu-CPS-1 chemical composition differed in sugar and uronic acid contents, and they were composed of nine constituent monosaccharides and one uronic acid with different ratios, with the average molecular weights of 1.02 × 10 3 , 1.12 × 10 3 and 1.33 × 10 3 kDa, respectively. They presented similar fourier transform infrared spectra, but different surface morphology, chain length and branching. Antioxidant assay showed that they all exhibited strong scavenging activity on ABTS + and hydroxyl radicals and moderate activity on DPPH radical. Glu-CPS-1 exhibited the highest antioxidant activity suggested culture conditions could regulate the bioactivity through influencing the structure and properties. These findings demonstrated the potential application of proper regulation of culture conditions in the development of polysaccharides with high antioxidant activity. [ABSTRACT FROM AUTHOR]

Cellulases like endoglucanase II (EGII) from Trichoderma reesei are the industrial enzymes responsible for breakdown of cellulosic materials. Due to its importance for production of eco-friendly commercial products such as alternative biofuels, industrial EGII production and optimization of its production conditions merit consideration. The gene responsible for EGII expression was designed and sub-cloned in to pET26b expression vector and transformed into BL21 (DE3) pLysS cells. Protein expression and purification was followed by a RSM design (20 experiments) to optimize the IPTG Concentration, post induction period and cell density (OD600). Thereafter, another RSM design (20 experiments) was performed to find and optimize the most important permeabilizing factors to achieve higher extracellular EGII expression. The EGII expression levels were assessed by Ghose method. The EGII gene was sub-cloned and protein expression and purification were successfully performed. The RSM experiments indicated that 0.331 mM for IPTG Concentration, 10.89 H for post induction period and 3.41 for cell density (OD600) were the optimum culture. Glycine (0.99%), Triton X-100 (0.73%) and CaCl2 (0.232) have been assigned as the most effective membrane permeabalizing factors. Optimization of culture medium components has led to a 3.06 fold increase in extracellular expression of EGII. RSM is an amenable method to optimize the expression of commercially significant enzymes. Our results indicated that optimization of IPTG concentration, post induction period and cell density along with glycine, Triton X-100 and Ca2+ concentration could lead to more cost effective industrial production of EGII. [ABSTRACT FROM AUTHOR]

Background: Aberrant DNA methylation patterns of genes required for development are common in in vitro produced embryos. In this regard, we previously identified altered DNA methylation patterns of in vivo developed blastocysts from embryos which spent different stages of development in vitro, indicating carryover effects of suboptimal culture conditions on epigenetic signatures of preimplantation embryos. However, epigenetic responses of in vivo originated embryos to suboptimal culture conditions are not fully understood. Therefore, here we investigated DNA methylation patterns of in vivo derived bovine embryos subjected to in vitro culture condition before, during or after major embryonic genome activation (EGA). For this, in vivo produced 2-, 8- and 16-cell stage embryos were cultured in vitro until the blastocyst stage and blastocysts were used for genome-wide DNA methylation analysis. Results: The 2- and 8-cell flushed embryo groups showed lower blastocyst rates compared to the 16-cell flush group. This was further accompanied by increased numbers of differentially methylated genomic regions (DMRs) in blastocysts of the 2- and 8-cell flush groups compared to the complete in vivo control ones. Moreover, 1623 genomic loci including imprinted genes were hypermethylated in blastocyst of 2-, 8- and 16-cell flushed groups, indicating the presence of genomic regions which are sensitive to the in vitro culture at any stage of embryonic development. Furthermore, hypermethylated genomic loci outnumbered hypomethylated ones in blastocysts of 2- and 16-cell flushed embryo groups, but the opposite occurred in the 8-cell group. Moreover, DMRs which were unique to blastocysts of the 2-cell flushed group and inversely correlated with corresponding mRNA expression levels were involved in plasma membrane lactate transport, amino acid transport and phosphorus metabolic processes, whereas DMRs which were specific to the 8-cell group and inversely correlated with corresponding mRNA expression levels were involved in several biological processes including regulation of fatty acids and steroid biosynthesis processes. Conclusion: In vivo embryos subjected to in vitro culture before and during major embryonic genome activation (EGA) are prone to changes in DNA methylation marks and exposure of in vivo embryos to in vitro culture during the time of EGA increased hypomethylated genomic loci in blastocysts. [ABSTRACT FROM AUTHOR]

A new halophilic actinomycetes strain wascharacterized and on the basis of physiological and 16S rDNA sequence, it was designated as Kocuria sp. strain rsk4. It was able to produce antibacterial metabolites and shows broad spectrum antibacterial activity against several gram-positive and gram-negative bacteria. Rsk4 showed highest antagonistic effect against multi drug resistant bacteria Staphylococcus aureus. Cultural and nutritional conditions of strain have been optimized under shake-flask conditions for the efficient production of antibacterial metabolites. Among the various factors, starch casein medium, 30ºC temperature, neutral pH, 5% salinity, 7 days incubation period and 180 rpm agitation was found to be optimal for antibacterial metabolite production. 1% starch, yeast extract, casein and magnesium were found to be best carbon source, nitrogen sources and mineral respectively for improved antibacterial activity. Among various combinations of nitrogen source, yeast extract and casein in equal ratio showed enhanced antibacterial activity. This is the first report on the optimization of antibacterial metabolites by Kocuria sp. The study suggests that the Kocuria sp. could scale up by traditional method for production of antibacterial metabolites effective against multiple drug resistant S. aureus. [ABSTRACT FROM AUTHOR]

The environmental conditions were optimized to produce the enniatin H, I, and MK1688 by Fusarium strain on cereal grain exhibiting anti-carcinogenic potential against MES-SA (human uterine sarcoma cell line), HCT15 (human colorectal carcinoma cancer cell line), and their multidrug resistance sublines. From the statistical optimization by response surface methodology, the optimal condition of independent variables affecting the response variables were 20.85 °C (temperature), 46.85% (w/w, initial moisture content), and 18.42 days (growth time) for ENN H; 23.31 °C, 44.15% (w/w) and 17.23 days for ENN I; 23.08 °C, 43.97% (w/w) and 17.06 days for ENN MK1688. In case of cytotoxic effects, ENNs significantly suppressed growth of cancer cell lines without multidrug resistance, and ENN I inhibited growth of cancer cell lines most strongly. These data will provide valuable point to produce the cyclic hexadepsipeptide exhibiting anti-carcinogenic potential from Fusarium strains., Competing Interests: Conflict of interestThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© The Korean Society of Food Science and Technology 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

A hiperidricidade, anteriormente chamada vitrificação, é considerada uma desordem fisiológica, bioquímica e morfológica decorrente do acúmulo anormal de água no interior das células e tecidos. As plantas cultivadas in vitro estão, indubitavelmente, sob contínua condição de estresse, os quais resultam em alterações metabólicas características do estresse oxidativo. Anatomicamente, plantas ou brotos afetados frequentemente apresentam-se inchados, com coloração verde claro, folhas translúcidas e com aparência de vidro, baixa relação número de células/área celular e hipolignificação. Alterações fisiológicas que ocorrem nas principais vias metabólicas, incluindo fotossíntese, respiração e transpiração, resultam em redução de eficiência dessas vias metabólicas. Os distúrbios morfológicos, fisiológicos e bioquímicos são desencadeados por fatores físicos, relacionados ao ambiente dos recipientes de cultivo e consistência do meio de cultura ou por fatores químicos como os componentes do meio de cultura, em especial dos reguladores de crescimento em altas concentrações. A hiperidricidade ocorre em vários níveis de severidade, chegando a resultar na perda irreversível da capacidade morfogênica e o estabelecimento de um estado neoplásico das células, no entanto, na maioria dos casos, a hiperidricidade é considerada reversível. Esta revisão foca o conhecimento atual sobre o fenômeno da hiperidricidade abordando aspectos morfológicos, fisiológicos, bioquímicos e a reversibilidade do processo.The hyperhydricity, formerly called vitrification, is considered a physiological, biochemistry and morfologic disorder due to abnormal accumulation of water inside the cells and tissues. Plants grown in vitro are undoubtedly under continuous stress condition which results in metabolic changes characteristic of oxidative stress. Anatomically plants or shoots affected often become swollen, with pale green, translucent sheets, glass-like, low relative number of cells / cell area and hipolignification. Physiological changes occur in major metabolic pathways including photosynthesis, respiration and transpiration resulting in reduced efficiency of these metabolic pathways. Morphological, physiological and biochemical disorders are triggered by physical factors related to the environment of cultivation vessels and consistency of the culture medium or by chemical factors such as culture medium components, especially the growth regulators in high concentrations. The hyperhydricity occurs at various levels of severity, reaching result in irreversible loss of morphogenic capacity and the establishment of a state of neoplastic cells, however, in most cases hyperhydricity is considered reversible. This review focuses on the current knowledge about the phenomenon of hyperhydricity addressing morphological, physiological, biochemical, and reversibility of the process.

The root of Angelica sinensis (Oliv.) Diels is a traditional Chinese medicine (TCM) widely used for the cardio-cerebrovascular, anti-inflammatory, and anti-cancer agents, etc. Currently, the root yield is mainly limited by narrow geographical distribution, early bolting and flowering (EBF), continuous cropping obstacle, and germplasm degeneration. While two ancillary strategies to produce bioactive metabolites by bioreactor and innovate new germplasms by genome editing have not been conducted. Here is reported embryogenic callus induction, histomorphological structure identification, suspension cell culture, bioactive metabolite determination, shoot bud and root differentiation, and plantlet acclimatization at different treatments such as explants, hormones, and culture conditions. The embryogenic calli can be effectively induced (99.0%) from the root of aseptic seedlings by a combination of 2.0 mg/L IBA and 0.2 mg/L KT, and embryogenic cells including: three-cell, globular-, heart-, torpedo-, and cotyledon-shaped embryos were observed. The maximum growth rate, bioactive metabolites contents (i.e. , ferulic acid, soluble sugar, flavonoids, and phenolic), and antioxidant capacities were observed at 9–12 d with inoculum density 40 g/L, 22 ℃ and 16/8 h light/dark. Robust plantlets can be obtained within 180 d after successive shoot bud and root differentiation as well as plantlet acclimation. Based on the strategies for the satisfaction of root demand, an effective and complete scheme is proposed for producing bioactive metabolites and regenerating plant of A. sinensis. [Display omitted] • Embryogenic calli are effectively induced (99.0%) from the root of aseptic seedlings. • Three-cell, globular-, heart-, torpedo- and cotyledon-shaped embryos are observed. • The highest metabolites production can be achieved from suspension cells at 9–12 d. • Plantlets are obtained by callus differentiation and plantlet acclimation within 180 d. • A scheme is proposed for producing bioactive metabolites and regenerating plants. [ABSTRACT FROM AUTHOR]

Received: 1/May/2007, Accepted: 5/Jul/2007The capacity of mesenchymal stem cells (MSCs) to differentiate amongskeletal cell lineages and to undergo extensive proliferation in vitro rendersthem an appropriate source for cell-replacement therapy to heal defects ofbone and cartilage tissue.It is argued that in bone and cartilage defects,MSCs would better be transplanted as fully-differentiated cells otherwise theymay produce non-specific cells in defect sites. This notion may emphasizethe importance of the studies considering in vitro bone and cartilagedifferentiation of MSCs. Indeed, the capacity of producing osteoblastic andchondrocytic cell lineage is among the earliest differentiation potentials ofMSCs, being reported at first isolation of the cells. Recent studies, howeverindicate that MSCs could differentiate into more cell lineages than expected.The present study provides evidence for, in vitro potential of MSCs todifferentiate into bone and cartilage cell lineages. As an introduction tothe differentiation, the characteristics of MSCs have been described andMSCs differentiation reviewed. The culture condition for bone and cartilagedifferentiation, molecular regulation of the differentiation, signaling pathwayinvolved during the differentiation, and the genes up-regulated upon boneand cartilage differentiation have also been described.

Secondary metabolite contents of W. somnifera varied remarkably between seasons and genotypes under ex vitro condition. In vitro studies provide an optimum culture condition for steady and quality production of bioactive chemicals throughout the year without involvement of environmental stresses. Mass production of micro-shoots and plantlets by exploring the organogenic totipotency of shoot tip explants (ex vitro and in vitro grown) considering two elite genotypes (Poshita and Jawahar 22) of W. somnifera, and assessment of their capability in production and accumulation of bioactive metabolites (total alkaloid and withanolides amount were quantified; withanolide A and withaferin A contents were estimated by High Performance Liquid Chromatography-HPLC). [ABSTRACT FROM AUTHOR]

Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors. Previously, we developed recombinant Escherichia coli coexpressing periplasmic organophosphorus hydrolase (OPH) and cytosolic chaperone GroEL-GroES (GroEL/ES). In present work, we investigated the effect of culture conditions on whole cell OPH activity. Especially, the whole cell OPH activity was significantly affected by the concentration of tetracycline that is an inducer for chaperone GroEL/ES. When cultured at 20°C for 31 h in M9 medium containing 1 mM IPTG, 50 ng/mL tetracycline, and 500 µM CoCl, the recombinant E. coli exhibited a specific whole cell OPH activity (U/OD) of ~0.55, which is 2.6-fold higher than that of recombinant E. coli cultured as previously described conditions. In addition, recombinant cells showed adequate storage stability for 1 week with 100% of original response. Finally, the improved activity and adequate stability in the whole cell biocatalyst will contribute to sensitivity, detection time, and stability of a whole cell-based biosensor for the detection of toxic organophosphates. [ABSTRACT FROM AUTHOR]

To evaluate the effect of atmospheric oxygen (O2) concentration on embryonic development, a controlled and randomized study using the sibling oocytes was carried out. A total of 147 patients were studied. Embryos were cultured in O2 concentration 20% versus 5% during the gamete, zygote, and first 3 days. The mean cell numbers of embryo (7.69 ± 1.91 vs 7.20 ± 1.82, P = .011) and rate of clinically useable embryos (81.62% vs 77.22%, P = .017) were significantly higher in 5% O2 than in 20% O2. There was no difference in the zygote developmental stage, day 2, day 4, and blastocyst stage. The quality of blastocyst (both inner cell mass and trophectoderm) showed no difference. Also, there was no increase in embryos fragmentation and uneven cells in 20% O2 culture condition. In conclusion, 20% O2 reduced the mean cell numbers of embryo and the number of clinically useable embryos on day 3. However, there was no subsequent negative impact on development of day 4 and blastocyst stage. [ABSTRACT FROM AUTHOR]

Piriformospora indica , which is an endophytic fungus that grows on various media in the absence of a host, emits plant growth promoting volatile organic compounds (VOCs). Kaefer medium (KF) has been shown to be the most suitable medium for P. indica growth; however, different media may differentially affect fungal metabolism which may in turn influence the VOC profiles of P. indica . To date, how the VOCs emitted from P. indica cultured on different media affect plant growth has not been well characterized. Here, we show that poor nutrient medium (PNM) promoted the growth of P. indica more effectively than potato dextrose agar (PDA) or KF medium. By contrast, plant total biomass and root fresh weight were increased 1.8-fold and 2.1-fold, when co-cultivated with P. indica cultured on PDA medium in comparison with KF or PNM medium, respectively. Furthermore, sucrose in the plant culture medium downregulated the fold-induction ratio of the plant growth promoted by P. indica VOCs., Competing Interests: Conflict of interestThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 Japanese Society for Plant Biotechnology.)

In this study, we investigated the effect of cell density on the proliferation activity of human mesenchymal stem cells (MSCs) derived from adipose tissue (AT-MSCs) over time in culture. Passage #4 (P4) and #12 (P12) AT-MSCs from two donors were plated at a density of 200 (culture condition 1, CC1) or 5000 (culture condition 2, CC2) cells cm−2. After 7 days of incubation, P4 and P12 AT-MSCs cultured in CC1 were thin and spindle-shaped, whereas those cultured in CC2 had extensive cell-to-cell contacts and an expanded cell volume. In addition, P4 and P12 AT-MSCs in CC1 divided more than three times, while those in CC2 divided less than once on average. Flow cytometric analysis using 5(6)-carboxyfluorescein diacetate N-succinimidyl ester dye showed that the fluorescence intensity of AT-MSCs was lower in CC1 than in CC2. Furthermore, expression of proliferation-associated genes, such as CDC45L, CDC20A and KIF20A, in P4 AT-MSCs was higher in CC1 than in CC2, and this difference was also observed in P12 AT-MSCs. These data demonstrated that cell culture density affects the proliferation activity of MSCs, suggesting that it is feasible to design a strategy to prepare suitable MSCs using specific culture conditions. Copyright © 2016 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]