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Hesti Lina Wiraswati,1 Savira Ekawardhani,1 Enny Rohmawaty,1 Amila Laelalugina,2 Ade Zuhrotun,3 Rini Hendriani,4 Yoga Windhu Wardhana,5 Muhammad Begawan Bestari,6 Emmy Hermiyanti Sahirdjan,7 Sumartini Dewi7 1Department of Biomedical Sciences, Universitas Padjadjaran, Sumedang, West Java, Indonesia; 2Oncology and Stem Cell Working Group, Universitas Padjadjaran, Bandung, West Java, Indonesia; 3Department of Biological Pharmacy, Universitas Padjadjaran, Sumedang, West Java, Indonesia; 4Department of Pharmacology and Clinical Pharmacy, Universitas Padjadjaran, Sumedang, West Java, Indonesia; 5Study Center of Pharmaceutical Dosage Development, Department of Pharmaceutics and Pharmaceuticals Technology, Universitas Padjadjaran, Sumedang, West Java, Indonesia; 6Division Gastro Entero Hepatology, Department of Internal Medicine, Universitas Padjadjaran, Bandung, West Java, Indonesia; 7Department of Internal Medicine, Universitas Padjadjaran, Bandung, West Java, IndonesiaCorrespondence: Hesti Lina Wiraswati, Department of Biomedical Sciences, Faculty of Medicine, Universitas Padjadjaran, Sumedang, West Java, 45363, Indonesia, Tel +62-22-84288888, Fax +022-84288888, Email hesti.lina@unpad.ac.idPurpose: Physalis angulata Linn. (Ciplukan) is a plant widely used in traditional medicine in subtropical and tropical regions. Most studies focus on its antioxidant and anti-inflammatory activity. Many studies also reported its therapeutic potential for treating cancer, malaria, hepatitis, rheumatism, liver problems, and tumors, but few studies have reported its anti-fibrosis activity. Here, we aimed to investigate the potential of P. angulata as an antioxidant and anti-inflammatory that may be correlated with its anti-fibrosis action.Methods: In our study, we treated 3T3-L1 and TGF-β-induced 3T3-L1 cells with an ethanol extract of P. angulata. We then monitored the cell’s response, evaluated the antioxidant activity using an MTT assay, and observed the cells’ migration using the cell scratch assay. We used RT-PCR to determine the expression of HIF-1α and IL-6 on TGF-β-induced 3T3-L1 cells.Results: The ethanol extract of P. angulata showed antioxidant activity and promoted cell proliferation on 3T3-L1 cells. Interestingly, the extract inhibited the migration of TGF-β-induced 3T3-L1 cells. Further analysis revealed that the extract could inhibit HIF-1α expression and suppress IL-6 expression on TGF-β-induced 3T3-L1 cells.Conclusion: The ethanol extract of P. angulata showed antioxidant and anti-inflammation activities in 3T3-L1 cells. Both activities are associated with the antifibrotic activity of P. angulata’s ethanol extract.Keywords: TGF-β, ethanol extract, wound healing, fibroblast cell line

Agents that will accelerate wound healing maintain their clinical importance in all aspects. The aim of this study is to determine the antimicrobial activity of zinc oxide nanoparticles (ZnO NPs) ZnO nanoparticles obtained by green synthesis from Capparis spinosa L. extract and their effect on in vitro wound healing. ZnO NPs were synthesized and characterized using Capparis spinosa L. extract. ZnO NPs were tested against nine ATCC-coded pathogen strains to determine antimicrobial activity. The effects of different doses (0.0390625–20 µg/mL) of NPs on cell viability were determined by MTT assay. The effect of ZnO NPs doses (0.0390625 µg/mL, 0.078125 µg/mL, 0.15625 µg/mL, 0.3125 µg/mL, 0.625 µg/mL, 1.25 µg/mL) that increase proliferation and migration on wound healing was investigated in an in vitro wound experiment. Cell culture medium obtained from the in vitro wound assay was used for biochemical analysis, and plate alcohol-fixed cells were used for immunohistochemical staining. It was determined that NPs formed an inhibition zone against the tested Gram-positive bacteria. The ZnO NPs doses determined in the MTT test provided faster wound closure in in-vitro conditions compared to the DMSO group. Biochemical analyses showed that inflammation and oxidative status decreased, while antioxidant levels increased in ZnO NPs groups. Immunohistochemical analyses showed increased expression levels of Bek/FGFR2, IGF, and TGF-β associated with wound healing. The findings reveal the antimicrobial effect of ZnO nanoparticles obtained using Capparis spinosa L. extract in vitro and their potential applications in wound healing. [ABSTRACT FROM AUTHOR]

Background: Commiphoracaudatais a traditional medicinal plant which having various medicinal activities but the present research work was focussed on the wound healing activity. The leaves were extracted using continuous hot percolation method. Based on the phytochemical studies, the ethanolic extract was selected for the preparation of herbal tablet. Direct compression was done by making eight batches [F1-F8] of herbal tablets. The pre and post compression studies were investigated for the developed formulation to select optimized one. Aim and objective: The current investigation of the present research is to formulate and develop the herbal tablet from the ethanolic leaf extract of medicinal plant Commiphoracaudata(Wight &Arn.) Engl.and to evaluate the in vitro wound healing studies. Materials and methods: Petroleum ether, Chloroform, Acetone, Ethanol, HPMC, Carbopol, Starch, Lactose monohydrate, Crospovidone, Mannitol, Magnesium stearate, Disodium hydrogen phosphate, Potassium dihydrogen phosphate phosphate which are used for the extraction process and formulation and evaluation of the herbal tablet to analyse the wound healing studies. Result: From this formulation F7 shows the better results than the rest of other formulations and it was subjected for the fibroblast cell line studies(3T3-CCL92) resulted in a significant increase in the percentage from 34.7-53.29% and the dose-response curve produced to be linear which conform the wound healing activity. Then the formulation F7 were carried out for short term accelerated stability studies for 3 months which shows negligible changes. Conclusion: the herbal tablet used for the treatment of wound healing activity as a traditional system of medicine. [ABSTRACT FROM AUTHOR]

Background: Commiphoracaudatais a traditional medicinal plant which having various medicinal activities but the present research work was focussed on the wound healing activity. The leaves were extracted using continuous hot percolation method. Based on the phytochemical studies, the ethanolic extract was selected for the preparation of herbal tablet. Direct compression was done by making eight batches [F1-F8] of herbal tablets. The pre and post compression studies were investigated for the developed formulation to select optimized one. Aim and objective: The current investigation of the present research is to formulate and develop the herbal tablet from the ethanolic leaf extract of medicinal plant Commiphoracaudata(Wight &Arn.) Engl.and to evaluate the in vitro wound healing studies. Materials and methods: Petroleum ether, Chloroform, Acetone, Ethanol, HPMC, Carbopol, Starch, Lactose monohydrate, Crospovidone, Mannitol, Magnesium stearate, Disodium hydrogen phosphate, Potassium dihydrogen phosphate phosphate which are used for the extraction process and formulation and evaluation of the herbal tablet to analyse the wound healing studies. Result: From this formulation F7 shows the better results than the rest of other formulations and it was subjected for the fibroblast cell line studies(3T3-CCL92) resulted in a significant increase in the percentage from 34.7-53.29% and the dose-response curve produced to be linear which conform the wound healing activity. Then the formulation F7 were carried out for short term accelerated stability studies for 3 months which shows negligible changes. Conclusion: the herbal tablet used for the treatment of wound healing activity as a traditional system of medicine. [ABSTRACT FROM AUTHOR]

The yak lives in harsh alpine environments and the rumen plays a crucial role in the digestive system. Rumen-associated cells have unique adaptations and functions. The yak rumen fibroblast cell line (SV40T-YFB) was immortalized by introducing simian virus 40 large T antigen (SV40T) by lentivirus-mediated transfection. Further, we have reported the effects of lipopolysaccharide (LPS) of different concentrations on cell proliferation, extracellular matrix (ECM), and proinflammatory mediators in SV40T-YFB. The results showed that the immortalized yak rumen fibroblast cell lines were identified as fibroblasts that presented oval nuclei, a fusiform shape, and positive vimentin and SV40T staining after stable passage. Chromosome karyotype analysis showed diploid characteristics of yak (n = 60). LPS at different concentrations inhibited cell viability in a dose-dependent manner. SV40T-YFB treated with LPS increased mRNA expression levels of matrix metalloproteinases (MMP-2 and MMP-9), inflammatory cytokines (TNF-α, IL-1β, IL-6), and urokinase-type plasminogen activator system components (uPA, uPAR). LPS inhibits the expression of tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2), plasminogen activator inhibitor-2 (PAI-2), fibronectin (FN), anti-inflammatory factor IL-10, and collagen I (COL I) in SV40T-YFB. Overall, these results suggest that LPS inhibits cell proliferation and induces ECM degradation and inflammatory response in SV40T-YFB. [ABSTRACT FROM AUTHOR]

Background: 5-Fluorouracil (5-FU) is a routine anticancer agent for several cancer types, its short half-life requires continuous administration in high doses. This results in toxic side-effects to the normal tissues in the body. Vitamin C is an antioxidant which believed to impart cyto-protection to normal tissues in cancer patients treated with chemotherapeutic drugs. Aim of the Study: To assess the potential benefit of vitamin C administration 24 hours after 5-FU treatment compared to its immediate supplementation on normal fibroblast cell line. Materials and Methods: Fibroblast cell line (WI-38) was divided into 5 groups. Group I not treated (control group), group II treated with 5-FU only and group III received vitamin C only. Groups IV and V received 5-FU with vitamin C simultaneously and after 24 hours of 5-FU administration, respectively. The viability of cells in all groups was determined using MTT assay. MDA Quantification Assay was used to determine lipid peroxidation. Flow cytometry was used for evaluation of the cell cycle and cell death. All recorded data was statistically analyzed. Results: Cells in group IV seems like group II fewer and rounded while in group V some of them adherent spindle and the others rounded non-adherent. The least viability was noted in group II. MDA levels were significantly lower in groups III compared to all the other groups. Cell cycle analysis revealed the presence of statistically significant differences between groups in different phases of the cycle. Flowcytometry analysis showed the highest apoptosis in groups II and IV. Conclusion: Co-treatment of normal fibroblasts with vitamin C along with 5-FU reduces cell death and lipid peroxidation while enhancing viability and proliferation in treated fibroblasts, indicating healing and repair. Vitamin C administration 24 hours after 5-FU is more effective in reducing cell death and lipid peroxidation compared to its concomitant administration with 5-FU. [ABSTRACT FROM AUTHOR]

Introduction Collagen plays a vital role in maintaining the structural integrity of dentin, and its modification with bioactive compounds can enhance its mechanical properties and bonding capabilities. Aim This study aimed to evaluate the genotoxic effects of grape seed extract (GSE) and marine collagen peptide (MCP) on dental pulp-derived primary cells. Methodology Human dental pulp stem cells were isolated, cultivated, and then treated with GSE and marine collagen peptides. DNA fragmentation was assessed using DAPI (4',6-diamidino-2-phenylindole) staining. Statistical analysis was performed using SPSS version 20 (IBM Corp., Armonk, NY, USA). Results The results showed that GSE exhibited a minimum level of cell death compared to marine collagen peptides. The viable cell count increased steadily over three days in all groups, with the control group showing the highest number of viable cells. The differences in viable cell count among the groups were statistically significant. Conclusion This study suggests that GSE and marine collagen peptides are highly biocompatible with dental pulp cells and could be considered for further clinical studies., Competing Interests: Human subjects: All authors have confirmed that this study did not involve human participants or tissue. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, KLS et al.)

Isolation and culture of somatic cells from animals especially endangered species have raised great concerns as it is being an effective and convenient way to preserve genetic materials for future studies. As a species native to China, Chinese muntjac (Muntiacus reevesi) is listed as a beneficial species with economic and scientific research values. To our knowledge, however, there have been no published reports on somatic cell preservation of this species to date. To conserve biological resources for sustainability of Chinese muntjacs' genetic diversity, we established a fibroblast cell line from the postmortem ear skin of an adult male Chinese muntjac. The cultured cells were adherent to the plastic and showed an elongated, thin, and spindle-like shape. Moreover, they were FSP1- and VIM-positive characterizing them to be fibroblastic. No microorganisms (bacteria, fungi, or mycoplasmas) were detected throughout the whole study. Cell viability was high although it declined somehow after passaging. The population doubling time was 21.28 h according to the growth curve. Chromosome analysis revealed that the established fibroblast cell line contained 23 pairs of chromosomes, one pair of which was sex chromosomes (XY). Mitochondrial cytochrome C oxidase I gene of cultured cells shared 98.32% identity with those of Muntiacus reevesi registered in GenBank, which verified the cell line was derived from Muntiacus reevesi. In conclusion, we propagated and characterized fibroblast cells from a Chinese muntjac. We believe that this somatic cell line could facilitate animal cloning and breeding studies and become a useful in vitro model to address genetic questions. [ABSTRACT FROM AUTHOR]

Ethnic value of many known plants are underexploited for medicinal application besides their proven traditional qualities. One such plant known for wound healing is Tridax procumbens. This plant has wound healing property and is commercially unexploited. Silver nanoparticle (Ag‐NP) were synthesized using this plant extracts using different solvents (methanol, ethyl acetate and aqueous), which exhibit resonance at 426, 424 and 418 nm, respectively. This plant‐mediated Ag‐NPs have strong anti‐bactericidal activity against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Streptococcus pyogenes, Klebsiella pneumonia, Serratia marcescens and Bacillus subtilis with methanol extract. Further instance, elemental composition was confirmed by energy dispersive X‐ray analysis and particle size ranges were observed at 80–200 nm with spherical shape nanoparticles by scanning electron microscopy and transmission electron microscopy analysis. The biocompatibility of Ag‐NPs was assessed using fibroblast cell line (L929) by MTT assay with 109.35 µg IC50 value. The oxygen plasma treated and non‐treated bamboo spunlaced nonwoven fabrics were coated with the Ag‐NPs by exhaust method. Contact angle and water retention revealed significant difference in absorption ability of plasma treated fabric. Field emission scanning electron microscopy revealed the presence of Ag‐NPs in plasma coated fabrics. The fabricated cloth was studied for anti‐microbial and microbial penetration ability. [ABSTRACT FROM AUTHOR]

The study examines the phenolic compounds in hydromethanolic extracts of Salix alba (L.) leaves and bark as well as their antioxidant activity and cytotoxic potential. UPLC-PDA-Q/TOF-MS analysis showed a total of 29 phenolic compounds in leaves and 34 in bark. Total phenolic compound content was 5575.96 mg/100 g of dry weight (DW) in leaves and 2330.31 mg/100 g DW in bark. The compounds were identified as derivatives of phenolic acids (seven in leaves and five in bark), flavanols and procyanidins (eight in leaves and 26 in bark) and flavonols (14 in leaves and three in bark). Both extracts exhibited strong antioxidant potential, assessed by radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), but the bark extract was even stronger than the ascorbic acid used as a standard. The cytotoxicity of both extracts was evaluated against human skin fibroblasts and human epidermal keratinocytes cell lines using the Presto Blue cell viability assay. The keratinocytes were more resistant to tested extracts than fibroblasts. The leaf and bark extracts at concentrations which exhibited antioxidant activity were also not toxic against the keratinocyte cell line. Thus, S. alba extracts, especially the leaf extract, offer promise as a nontoxic natural antioxidant, in cosmetic products or herbal medicines, and as a source of bioactive secondary metabolites.

Caspian horse, a rare horse breed found in 1965 by Louise Firouz in northern Iran, is a small horse which is reported to be in danger of extinction in its original homeland. There seems to be a great need to prevent extinction of this valuable horse. In this study, 51 fibroblast cell lines from Caspian horse ear marginal tissue were successfully established by sampling 60 horses using primary explant technique. Cells were authenticated and growth curve was plotted. According to results obtained, population doubling time (PDT) was calculated 23 ± 0.5 h for all cell lines. Multiplex polymerase chain reaction (multiplex PCR) revealed that cell lines had no cross-contamination with other species. Bacteria, fungi, and mycoplasma contamination were checked using standard methods such as PCR, direct culture, and Hoechst staining. In addition to providing a valuable source for genomic, postgenomic, and somatic cloning researches, the established cell lines would preserve Caspian horse genetic resources. It will also create an accessible database for researchers. [ABSTRACT FROM AUTHOR]

In this study, in vitro and in vivo methods were used to evaluate the cytotoxicity and genotoxicity properties of silver nanoparticles (Ag-NPs) made from a crude ethanolic extract of Salacia chinensis. The test Ag-NPs had no cytotoxicity on the fibroblast cell line at a concentration of 100 μg/mL, according to the MTT assay results. The Chinese hamster ovary (CHO) cell line treated with varied concentrations of test Ag-NPs, with a maximum concentration of 200 μg/mL, did not exhibit any appreciable genotoxic activity, either by comparing the results with positive controls of genotoxicity caused by Methyl methane sulfonate and Benzo (a) pyrene at the concentration of 20 μg/mL, the lack of genotoxicity was established. An in vivo study in Swiss albino mice using various concentrations (250, 500, and 1000 mg/kg) of test Ag-NPs, which were compared with positive controls, further confirmed this in vitro result pattern. Contrary to the genotoxicity caused by the positive control, mouse bone marrow micronucleus testing findings revealed the absence of genotoxicity. These findings imply that at the measured doses, the Ag-NPs produced from the crude ethanolic extract of Salacia chinensis do not exhibit any cytotoxicity or genotoxicity. • The AgNPs not causing cytotoxicity on fibroblast cellline at100 μg/mL concentration. • No chromosomal aberration found in CHO cellline at 200 μg mL−1 dosage of AgNPs. • The AgNPs not inducing any genotoxicity on both in-vitro and in-vivo studies. • The animal study proved that absence of genotoxicity on bone marrow micronucleus test. • S. chinensis synthesized AgNPs not causing cytotoxicity and genotoxicity. [ABSTRACT FROM AUTHOR]

Polygonum hydropiper is used as anti-cancer and anti-rheumatic agent in folk medicine. This study was designed to investigate the anti-angiogenic, anti-tumor, and cytotoxic potentials of different solvent extracts and isolated saponins. Samples were analyzed using GC, Gas Chromatography-Mass Spectrometry (GC-MS) to identify major and bioactive compounds. Quantitation of antiangiogenesis for the plant's samples including methanolic extract (Ph.Cr), its subsequent fractions; n-hexane (Ph.Hex), chloroform (Ph.Chf), ethyl acetate (Ph.EtAc), n-Butanol (Ph.Bt), aqueous (Ph.Aq), saponins (Ph.Sp) were performed using the chick embryo chorioallantoic membrane (CAM) assay. Potato disc anti-tumor assay was performed on Agrobacterium tumefaciens containing tumor inducing plasmid. Cytotoxicity was performed against Artemia salina and mouse embryonic fibroblast NIH/3T3 cell line following contact toxicity and MTT cells viability assays, respectively. The GC-MS analysis of Ph.Cr, Ph.Hex, Ph.Chf, Ph.Bt, and Ph.EtAc identified 126, 124, 153, 131, and 164 compounds, respectively. In anti-angiogenic assay, Ph.Chf, Ph.Sp, Ph.EtAc, and Ph.Cr exhibited highest activity with IC50 of 28.65, 19.21, 88.75, and 461.53 μg/ml, respectively. In anti-tumor assay, Ph.Sp, Ph.Chf, Ph.EtAc, and Ph.Cr were most potent with IC50 of 18.39, 73.81, 217.19, and 342.53 μg/ml, respectively. In MTT cells viability assay, Ph.Chf, Ph.EtAc, Ph.Sp were most active causing 79.00, 72.50, and 71.50% cytotoxicity, respectively, at 1000 μg/ml with the LD50 of 140, 160, and 175 μg/ml, respectively. In overall study, Ph.Chf and Ph.Sp have shown overwhelming results which signifies their potentials as sources of therapeutic agents against cancer. [ABSTRACT FROM AUTHOR]

This study aimed to establish the fibroblast cell bank of Qinghai fine-wool sheep in the form of somatic cells, and to study its biological characteristics, to help for long-term conservation and sustainable use of this genetic resource. Ear marginal tissue of the Qinghai Fine-wool sheep was used to establish a fibroblast cell line by direct culture of explants and cryopreservation techniques, and the quality of the cell line was assessed. This study not only shows the effective preservation of the Qinghai Fine-wool sheep germplasm at the cellular level, but also provides a available experimental resource for fields including cell biology, genomics, post-genomics, genetic engineering and embryo engineering. [ABSTRACT FROM AUTHOR]

Abstract: Establishment of fibroblast cell lines of endangered pig breeds and research on the gene functions based on the cells made a significant contribution to the conservation and utilization of genetic resources. The Wuzhishan miniature pig ear marginal tissue fibroblast cell line (WPF22) from 22 samples, stocking 87 cryogenically-preserved vials, was successfully established by using primary explants technique and cell cryopreservation techniques. WPF22 cells were adherent, with a population doubling time of 30.2h. Chromosome karyotyping and G-banding analysis showed that >90.2% of cells were diploid (2n =38) prior to the 4th generation. Neither microbial contamination nor cross-contamination was detected by isoenzyme analyses. Cell viability was 97.8% before cryopreservation and 94.9% after recovery. To determine cell permeability, intracellular path and stability of exogenous proteins during the transduction, six fluorescent protein genes were transferred into fibroblasts by lipofectamine-mediated method. The transfection efficiency of six fluorescent protein genes fluctuated between 8.1% and 42.6%. ECFP and DsRed were mostly shown in cytoplasmic in dots around the nucleus, and EYFP and EGFP had a slightly stronger expression in the nucleus than in the cytoplasm, but without expression in some vacuoles. Every index of the WPF22 cell line meets all the standard quality controls of American type Culture Collection (ATCC). This research thus does not only preserve important genetic resources of Wuzhishan miniature pig at the cell level, but also serve as a valuable resource for genome, postgenome and somacloning research. [Copyright &y& Elsevier]

Establishment of fibroblast cell lines of endangered goat breeds and research on the gene or protein functions based on the cells made a significant contribution to the conservation and utilization of genetic resources. In this study, a fibroblast cell line of Angora goat (AGF31) from 31 samples, frozen in 162 cryogenically-preserved vials with (2-3) x 106 cells each, was successfully established by using explants culture and cryopreservation techniques. The cells cultured in vitro showed typical morphology of fibroblasts, with a population doubling time of 34.3h; The average viability of the cells was 97.6% before freezing and 95.9% after thawing; Chromosome karyotyping and G-banding analysis showed that >90.8% cells were diploid (2n=60) prior to the 4th generation; Neither microbial contamination nor cross-contamination was detected by isoenzyme analyses; To determine cell permeability, intracellular path and stability of exogenous proteins during the transduction, six fluorescent protein genes were transferred into fibroblasts by lipofectamine-mediated method. The transfection efficiency of six fluorescent protein genes fluctuated between 13.4% and 60.8%. ECFP and DsRed were mostly shown in cytoplasmic in dots around the nucleus, and EYFP and EGFP had a slightly stronger expression in the nucleus than in the cytoplasm. Angora goat fibroblasts still kept stable genetic properties and normal biological characteristics in vitro, and every index met the standard quality control of American Type Culture Collection. [ABSTRACT FROM AUTHOR]

In this study, a full-length enriched cDNA library was successfully constructed from Bengal tiger, Panthera tigris tigris, the most well-known wild Animal. Total RNA was extracted from cultured Bengal tiger fibroblasts in vitro. The titers of primary and amplified libraries were 1.28 × 106 pfu/mL and 1.56 × 109 pfu/mL respectively. The percentage of recombinants from unamplified library was 90.2% and average length of exogenous inserts was 0.98 kb. A total of 212 individual ESTs with sizes ranging from 356 to 1108 bps were then analyzed. The BLASTX score revealed that 48.1% of the sequences were classified as a strong match, 45.3% as nominal and 6.6% as a weak match. Among the ESTs with known putative function, 26.4% ESTs were found to be related to all kinds of metabolisms, 19.3% ESTs to information storage and processing, 11.3% ESTs to posttranslational modification, protein turnover, chaperones, 11.3% ESTs to transport, 9.9% ESTs to signal transducer/cell communication, 9.0% ESTs to structure protein, 3.8% ESTs to cell cycle, and only 6.6% ESTs classified as novel genes. By EST sequencing, a full-length gene coding ferritin was identified and characterized. The recombinant plasmid pET32a-TAT-Ferritin was constructed, coded for the TAT-Ferritin fusion protein with two 6× His-tags in N and C-terminal. After BCA assay, the concentration of soluble Trx-TAT-Ferritin recombinant protein was 2.32 ± 0.12 mg/mL. These results demonstrated that the reliability and representativeness of the cDNA library attained to the requirements of a standard cDNA library. This library provided a useful platform for the functional genome and transcriptome research of Bengal tigers. [ABSTRACT FROM AUTHOR]

Establishment of fibroblast cell lines of endangered goat breeds and research on the gene or protein functions based on the cells made a significant contribution to the conservation and utilization of genetic resources. In this study, a fibroblast cell line of Liaoning cashmere goat, frozen in 174 cryovials with 5 × 10 cells each, was successfully established from 60 goats ear marginal tissues using explant culture and cryopreservation techniques. Biological analysis of in vitro cultured cell line showed that, the cells were morphologically consistent with fibroblasts; the average viability of the cells was 94.9 % before freezing and 90.1 % after thawing; the growth process of cells was consisted of a lag phase, a logarithmic phase and a plateau phase; cell population doubling time was 65.5 h; more than 90 % of cells were diploid prior to the 6th generation; Neither microbial contamination nor cross-contamination was detected. To determine cell permeability, intracellular path and stability of exogenous proteins during the transduction, a TAT protein transduction domain was fused to the C-terminus of enhanced green fluorescent protein, the established fibroblast cell line was treated with the purified exogenous proteins at various concentrations by adding them to the cell culture media for 1-24 h and assayed cell morphology and protein presence, it was found that the purified exogenous proteins readily entered cells at a concentration of 0.1 mg/ml within 1.5 h and some of them could translocate into nucleus, moreover, the exogenous proteins appeared to be stable inside cells for up to 24 h. [ABSTRACT FROM AUTHOR]

The Yangtze finless porpoise ( Neophocaena phocaenoides asiaeorientalis), as the sole freshwater subspecies of N. phocaenoides, is endemic to the Yangtze River and its adjacent lakes. Its population has declined significantly over recent decades. In this study, we established a skin-derived finite fibroblast cell line of the Yangtze finless porpoise, named YFP-SF1, using primary cell culture methods, and an immortalized cell line, T-YFP-SF1, through co-transfection (GFP and SV40 T antigens) techniques. YFP-SF1 proliferated continuously with a minimum population doubling time of 31 h and exhibited age-dependent changes in growth rate. T-YFP-SF1 cells exhibited fibroblast morphology and were characterized by a shorter doubling time, higher attachment efficiencies, colony formation at a low seeding density, and growth in low serum concentrations. Anchorage independence and foci formation in the cell monolayer were observed from passage 36. The chromosome number of YFP-SF1 and T-YFP-SF1 remained stable at 2 n = 44 in the early passages, and the viability of thawed cells remained above 90% after cryopreservation in liquid nitrogen. Taken together, we have established fibroblast cell lines of Yangtze finless porpoise for the first time, which might assist as an in vitro model for this endangered mammal. [ABSTRACT FROM AUTHOR]

Abstract: The synthesis of a novel series of aminoquinoline derivatives 1a–p and their antiproliferative activities against A375 human melanoma cell line were described. Most compounds showed superior antiproliferative activities to Sorafenib as a reference compound. Among them, quinolinyloxymethylphenyl compounds 1k and 1l exhibited potent activities (IC50 =0.77 and 0.79μM, respectively) and excellent selectivity against melanoma and fibroblast cell lines. [Copyright &y& Elsevier]

A Texel sheep ear marginal tissue fibroblast cell line (named TSF19) was successfully established by using a primary explant technique and cell cryoconservation technology. TSF19 cells were adherent, with a population doubling time of 24.9 h. Chromosome analysis showed that >90% of cells were diploid prior to cell passage 4. Isoenzyme analyses of lactate dehydrogenase and malate dehydrogenase showed that the TSF19 cells had no cross-contamination with other species. Tests for cell line contamination with bacteria, fungi, or mycoplasmas were also negative. Plasmids encoding the fluorescent proteins pEGFP-N3, pECFP-N1, pDsRed1-N1, and pEYFP-N1 were transfected into cells to study exogenous gene expression in the cells. The plasmid transfection efficiency was between 21.8% and 46.5%. This newly established cell line will not only preserve the genetic resources of the important Texel sheep at the cell level but will also provide a valuable resource for genomic, postgenomic, somatic cloning research. [ABSTRACT FROM AUTHOR]

Hyperbaric oxygen (HBO) therapy is widely used to treat problem wounds associated with pathologic conditions compromising blood supply and tissue oxygenation because increased tissue oxygen levels enhance collagen synthesis, cell proliferation, and angiogenesis. However, little is known about the dose of hyperoxia needed to achieve optimal therapeutic effects. Moreover, HBO, by enhancing the production of reactive oxygen species (ROS), may also exert cytotoxic effects. In vitro models are simplified systems that may aid the development of treatment protocols with HBO. Hence, we have investigated the effects of HBO on the growth and ROS production of the 3T3/J2 fibroblast cell line in relation to the pressure and the duration of exposure.3T3/J2 fibroblasts were plated (5 × 103 cells/cm2) on six-well microtiter plates in phosphate buffered saline (PBS), put in a compression chamber, and exposed to 100% oxygen at a pressure of 1.0 or 2.5 atmosphere absolute (ATA) for 15, 30, 60, or 120 minutes. Then the cells were incubated in Dulbecco's modified minimum essential medium (DMEM) for 24, 48, or 72 hours, and at the end of the post-HBO incubation period, their number was determined. In other experiments, cells were detached just after HBO exposure, seeded on 60 mm Petri dishes, and cultured for 10 days in DMEM, and the colony forming units were counted. The effects of HBO exposure (2.5 ATA) on the apoptotic rate of cultured cells were investigated by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and enzyme-linked immunosorbent assays. To measure ROS production, 60 minutes before HBO exposure, 2′,7′-dichlorofluorescin (DCF) diacetate (200 nmol/mL) was added to PBS, and after HBO exposure (2.5 ATA), cells were lysated, and fluorescence-emission intensity was measured and converted to μmol DCF/μg protein.At 1.0 ATA, all HBO exposures increased the proliferation rate of cultured fibroblasts and their clonal growth efficiency. At 2.5 ATA, 15-minute exposure to HBO was ineffective, whereas 30- and 60-minute exposures raised the proliferation rate and clonal growth efficiency. Conversely, a 120-minute exposure significantly decreased these parameters compared with control cultures. The exposure of cells to HBO at 2.5 ATA for 120 minutes raised the apoptotic rate of cultured fibroblasts, whereas shorter exposure times were ineffective. All exposure periods to HBO at 2.5 ATA enhanced ROS production from cultured fibroblasts.Collectively, our findings allow us to conclude that (1) all of the exposure periods to HBO at 1.0 ATA or 30- and 60-minute periods at 2.5 ATA enhance cell growth, (2) 120-minute exposure to HBO at 2.5 ATA exerts a marked proapoptotic effect, and (3) no evident relationships occur between the effects of HBO on cell growth and ROS production. [ABSTRACT FROM AUTHOR]

A primary cell line (designated as CCf) derived from caudal fin tissue of channel catfish, Ictalurus punctatus, was developed using explant techniques. The cell line grew fastest in media supplied with FBS and channel catfish serum. The duplication time of the cell line under optimal conditions was ∼56 h at a plating density of 1.1 × 105 cells/ml. The cell line has been propagated continuously for 25 passages (1:4 dilution per passage), cryopreserved, and recovered successfully at different passages. The cultured cells had fibroblastic morphology, and synthesized fibronectin and Type I and III collagens in the cytoplasm. The cell line maintained the normal diploid chromosome number (58) of channel catfish throughout the experiment. Nucleolus organizer regions were located on the short arms of a pair of medium-sized submetacentrics, which is typical for channel catfish. This study provides a method for acquiring a cell line from juvenile catfish without sacrifice, and is especially useful for early screening of valuable fishes. [ABSTRACT FROM AUTHOR]