Search

Skeletal muscle satellite cells (SMSCs) play an important role in regulating muscle growth and regeneration. Chromatin accessibility allows physical interactions that synergistically regulate gene expression through enhancers, promoters, insulators, and chromatin binding factors. However, the chromatin accessibility altas and its regulatory role in ovine myoblast differentiation is still unclear. Therefore, ATAC-seq and RNA-seq analysis were performed on ovine SMSCs at the proliferation stage (SCG) and differentiation stage (SCD). 17,460 DARs (differential accessibility regions) and 3732 DEGs (differentially expressed genes) were identified. Based on joint analysis of ATAC-seq and RNA-seq, we revealed that PI3K-Akt, TGF-β and other signaling pathways regulated SMSCs differentiation. We identified two novel candidate genes, FZD5 and MAP2K6, which may affect the proliferation and differentiation of SMSCs. Our data identify potential cis regulatory elements of ovine SMSCs. This study can provide a reference for exploring the mechanisms of the differentiation and regeneration of SMSCs in the future., Competing Interests: Declaration of competing interest No conflict of interest for every authors., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Although melatonin has been shown to exhibit a wide variety of biological functions, its effects on promoting differentiation of neural cells remain unknown. Wnt signaling mediates major developmental processes during embryogenesis and regulates maintenance, self-renewal, and differentiation of adult mammalian stem cells. However, the role of the noncanonical Wnt pathway during neurogenesis remains poorly understood. In this study, the amniotic epithelial cells ( AECs) were isolated from bovine amnion and incubated with various melatonin concentrations (0.01, 0.1, 1, 10, or 100 μm) and 5 × 10(-5) m all-trans retinoic acid (RA) for screening optimum culture medium of neural differentiation, compared with each groups, 1 μm melatonin and 5 × 10(-5) m RA were selected to induce neural differentiation of AECs, and then siMT1, siMT2, oWnt-4, and siWnt-4 were expressed in AECs to research role of these genes in neural differentiation. Efficiency of neural differentiation was evaluated after expressed above genes using flow cytometry. Cell function of neural cells was demonstrated in vivo using spinal cord injury model after cell transplantation, and damage repair of spinal cord was assessed using cell tracking and Basso, Beattie, Bresnahan Locomotor Rating Scale scores. Results demonstrated that melatonin stimulated melatonin receptor 1, which subsequently increased bovine amniotic epithelial cell vitality and promoted differentiation into neural cells. This took place through cooperation with Wnt-4. Additionally, following cotreatment with melatonin and Wnt-4, neurogenesis gene expression was significantly altered. Furthermore, single inhibition of melatonin receptor 1 or Wnt-4 expression decreased expression of neurogenesis-related genes, and bovine amniotic epithelial cell-derived neural cells were successfully colonized into injured spinal cord, which suggested participation in tissue repair., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Bovine liver-derived mesenchymal stem cells (bLMSCs) were isolated from the liver tissue of 4-6 months old fetal calf, and then characterized by immunofluorescence and RT-PCR. We found that primary bLMSCs could be subcultured to 44 passages, the total culture time in vitro was 192 days. The results of surface antigen detection showed that bBMSCs expressed CD29, CD44, CD73, CD90, CD106 and CD166 but not expressed endothelial cells and hematopoietic cells specific marker CD34, CD45 and BLA-DR. The results of growth kinetics, colony-forming cell assay and cell cycle analysis indicated that the fetal bovine LMSCs had good proliferation ability in vitro. The cells from passages 7 were successfully induced to differentiate into osteoblasts, adipocytes and chondrocytes. The results indicate the potential for multi-lineage differentiation of bLMSCs that may represent an ideal candidate for cellular transplantation therapy.

The DMSCs (dermal mesenchymal stem cells) are multipotent stem cells, which can differentiate in vitro into many cell types. Much work has been done on DMSCs from humans, mice, rabbits and other mammals, but the related literature has not been published about these cells in cattle. In this study, we isolated and established the DMSC lines from cattle, thereby initiating further research on these cells, such as growth kinetics, detection of special surface antigen and RT-PCR (reverse transcription-PCR) assays to identify the biological characterization of the cell line. Furthermore, the DMSCs are induced to differentiate into adipocytes, osteoblasts and neural cells in vitro Our results suggest that DMSCs isolated from cattle possess similar biological characteristics with those from other species. Their multi-lineage differentiation capabilities herald a probable application model in tissue engineering and induced pluripotent stem cells., (© 2014 The Author(s).)

The use of amnion and amniotic fluid (AF) are abundant sources of mesenchymal stem cells (MSCs) that can be harvested at low cost and do not pose ethical conflicts. In human and veterinary research, stem cells derived from these tissues are promising candidates for disease treatment, specifically for their plasticity, their reduced immunogenicity, and high anti-inflammatory potential. This work aimed to obtain and characterize bovine amniotic fluid mesenchymal stem cells (AFMSC). The bovine AF from the amniotic cavity of pregnant gilts in the early stages of gestation (3- and 4-m-old bovine embryos) was collected. AFMSCs exhibit a fibroblastic-like morphology only starting from the fourth passage, being heterogeneous during the primary culture. Immunofluorescence results showed that AFMSCs were positive for β-integrin, CD44, CD73 and CD166, but negative for CD34, CD45. Meanwhile, AFMSCs expressed ES cell markers, such as Oct4, and when appropriately induced, are capable of differentiating into ectodermal and mesodermal lineages. This study reinforces the emerging importance of these cells as ideal tools in veterinary medicine; future studies aimed at a deeper evaluation of their immunological properties will allow a better understanding of their role in cellular therapy.

Dermis-derived mesenchymal stem/progenitor cells (DMS/PCs) were a multipotential stem cell population, which has potential applications in the tissue damage repair and skin transplant. Although a large number of studies deal with the human DMS/PCs self-renewal and regulation, however, the study of livestock-derived DMS/PCs has rarely been reported. Here, sheep DMS/PCs were isolated from one-month-old sheep embryos and studied at the cellular and molecular level. And then the DMS/PCs biological characteristics were analysed by RT-PCR and immunofluorescence. Experimental results showed that DMS/PCs could be expanded for 48 passages and the cells viability and hereditary character were steady. In addition, the DMS/PCs maker β -integrin, CD71, CD44, and CD73 were expressed positively through RT-PCR and immunofluorescence. Passage 3 DMS/PCs were successfully induced to differentiate into adipocytes, osteoblasts, chondrocytes, and neurocytes, respectively. The above results suggest that DMS/PCs not only have strong self-renewal capacity but also have the potential to differentiate into adipocytes, osteoblasts, chondrocytes, and neurocytes. The study provides theoretical basis and experimental evidence for potential clinical application.

Objective: To isolate and culture skeletal muscle satellite cells of Luxi cattle embryo and study its biological characteristics., Methods: The skeletal muscles were taken from the limbs of Luxi cattle embryo of 30-50 d old to isolate and culture the skeletal muscle satellite cells by differential adherence method and co-digestion of type I collagenase and trypsin. Satellite cell surface markers, desmin, MyoD, c-Met, Myf5 and pax7 were detected by immunocytochemistry and RT-PCR, and its biological characteristics were researched., Results: The satellite cells were cultured to over passage 15. Cell viability was (97.90 ± 0.96)% after cryopreservation. The immunocytochemical staining showed that the specific surface antigen markers desmin and MyoD were positive, and RT-PCR also indicated the positive expressions of desmin, Myf5, c-Met and pax7. Colony formation was (56.39 ± 1.41)%. The karyotype analysis demonstrated that the isolated satellite cells were all derived from normal cattle embryos. Bovine satellite cells were successfully induced into osteoblasts and neuron-like cells by various inductors, and were positively expressed after alizarin red and toluidine blue staining. The osteoblast specific genes osteopontin and type I collagen and the nerve cell specific genes MAP-2 and nestin were positive by RT-PCR., Conclusion: The skeletal muscle satellite cells of the Luxi cattle embryo are successfully isolated and cultured in vitro , They have the ability of being differentiated into osteoblasts and neuron-like cells.

Mesenchymal stem/progenitor cells derived from Wharton's jelly of the umbilical cord (UCMSCs/UCMPCs) are multipotent, and can be differentiated in vitro into many cell types. Much work has been done on UCMSCs/UCMPCs from humans, mice, rabbits, and other mammals, but the relatively little literature has been published about these cells in chickens. In our work, we isolated USMSCs/USMPCs from chicken embryos. We characterized the isolated cells using immunofluorescence and reverse transcription polymerase chain reaction techniques. Primary UCMSCs/UCMPCs were subcultured to passage 30 and growth curves for each passage determined. The growth curves at different passages were all typically sigmoidal. Isolated UCMSCs/UCMPCs were induced to differentiate into adipocytes, osteoblasts, myocardial cells, and neural cells, and we were able to detect characteristic CD44, CD29, CD73, and CD71 cell surface markers. Our results suggest that UCMSCs/UCMPCs isolated from chickens possess similar biological characteristics to those from other species. Their multi-lineage differentiation capabilities herald a probable application for cellular transplant therapy in tissue engineering.

The bone marrow mesenchymal stem cells (MSCs) are multipotent stem cells which can differentiate into mesenchymal cells in vitro. In this study, MSCs in duck were isolated from bone marrow by density gradient centrifuge separation, purified and expanded in the medium. The primary MSCs were expanded for passages. The different-passage MSCs were induced to differentiate into osteoblasts and neuron-like cells. Karyotype analysis indicated that MSCs kept diploid condition and the hereditary feature was stable. The different-passage MSCs expressed CD44, ICAM- and SSEA-4, but not CD34, CD45 and SSEA-when detected by immunofluorescence staining. There was no significant difference among the positive rates of passages 2, 6 and 8 (P > 0.05), but a significant difference existed among those of passages 2, 6, 8 and 11 (P < 0.05). After the osteogenic inducement was added, the induced different-passage MSCs expressed high-level alkaline phosphatase (ALP), and are positive for tetracycline staining, Alizarin Red staining and Von Kossa staining. After the neural inducement was added, about 70% cells exhibited typical neuron-like phenotype, the induced different-passage MSCs expressed Nestin, neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) when detected by immunofluorescence staining. There was no significant difference among the positive rates of passages 3, 4 and 6 (P>0.05), but a significant difference existed among those of passages 3, 4, 6 and 8 (P<0.05). These results suggest that MSCs in duck were capable of differentiating into osteoblasts and neuron-like cells in vitro.

Embryonic stem (ES) cells have the unique capacity to proliferate extensively and maintain the potential to differentiate into advanced derivatives of all three primary germ layers. ES cell lines can also be generated from human blastocyst embryos and are considered promising donor sources for cell transplantation therapies for diseases such as juvenile diabetes, Parkinson's disease, and heart failure. However, as for organ transplants, tissue rejection remains a significant concern for ES cell transplantation. Another concern is the use of human embryos. One possible means to avoid these issues is by reprogramming the nuclei of differentiated cells to ES cell-like, pluripotent cells. This review discusses the potential of these strategies to generate tailor-made pluripotent stem cells and the role of transcription factors in the reprogramming process.

Melatonin can modulate neural stem cell ( NSC) functions such as proliferation and differentiation into NSC-derived pluripotent stem cells (N- iPS) in brain tissue, but the effect and mechanism underlying this are unclear. Thus, we studied how primary cultured bovine NSCs isolated from the retinal neural layer could transform into N- iPS cell. NSCs were exposed to 0.01, 0.1, 1, 10, or 100 μ m melatonin, and cell viability studies indicated that 10 μ m melatonin can significantly increase cell viability and promote cell proliferation in NSCs in vitro. Thus, 10 μ m melatonin was used to study miR-302/367-mediated cell reprogramming of NSCs. We noted that this concentration of melatonin increased reprogramming efficiency of N- iPS cell generation from primary cultured bovine NSCs and that this was mediated by downregulation of apoptosis-related genes p53 and p21. Then, N- iPS cells were treated with 1, 10, 100, or 500 μ m melatonin, and N- iPS (M-N- iPS) cell proliferation was measured. We noted that 100 μ m melatonin increased proliferation of N- iPS cells via increased phosphorylation of intracellular ERK1/2 via activation of its pathway in M-N- iPS via melatonin receptors 1 ( MT1). Finally, we verified that N- iPS cells and M-N- iPS cells are similar to typical embryonic stem cells including the expression of pluripotency markers (Oct4 and Nanog), the ability to form teratomas in vivo , and the capacity to differentiate into all three embryonic germ layers. [ABSTRACT FROM AUTHOR]