ABSTRACT
Introduction: The transdifferentiation potential of mesenchymal stem cells (MSCs) is not limited to mesodermal derivatives but also to other cell types such as neuronal cells under appropriate cell culture conditions.Materials and methods: The present study characterizes the differentiation of Wharton's jelly (WJ) derived MSCs using neuronal conditioned medium (NCM) collected from cultured foetal brain cells.Results: After induction with NCM to neuronal stem cells (NSC), the WJ MSCs showed profound morphological changes showing multiple neurites extending from the cell body containing reminiscent of Nissl substance and single long axon-like processes. In RT PCR and immunocytochemistry, the induced neuronal cells showed a strong positive expression of neuronal markers Nestin, ß III tubulin and GFAP indicated that, the cells were reactive to NCM for differentiation. A significant (p < 0.01) increase in the level of secretome BDNF was observed in NCM suggests that the BDNF could play a key role in the transdifferentiation of WJMSCs to NSCs.Conclusion: These results support the potential of ovine MSCs isolated from umbilical cord WJ of abattoir derived foetuses to differentiate into neuronal stem cells and also provide a valuable experimental data for NSC transplant research in veterinary medicine.
Subject(s)
Brain-Derived Neurotrophic Factor/metabolism , Cell Transdifferentiation/physiology , Mesenchymal Stem Cells/physiology , Neural Stem Cells/physiology , Wharton Jelly , Animals , Culture Media, Conditioned , Embryo, Mammalian , Glial Fibrillary Acidic Protein/metabolism , Nestin/metabolism , Neural Stem Cells/ultrastructure , Neurites/ultrastructure , Sheep , Tubulin/metabolism , Umbilical Cord , Wharton Jelly/cytologyABSTRACT
The present study has evaluated the association of growth differentiation factor9 (GDF9) and bone morphogenetic protein15 (BMP15) mRNA expression in cumulus-oocyte complexes (COCs) of buffalo ovary during in vitro maturation (IVM). GDF9 and BMP15 are expressed specifically in mammalian oocytes and also participate in cumulus-oocyte crosstalk. Quantitative real-time polymerase chain reaction (qRT-PCR) technique was applied to investigate the relative abundance (RA) of GDF9 and BMP15 mRNA transcripts throughout the IVM process. Relative mRNA expression pattern of these specific genes were assessed in oocytes and cumulus cells at 0, 6, 12 and 24 h of in vitro culture. Our results revealed that RA of GDF9 during different hours of IVM showed significant reduction between 0 h and 24 h of maturation in oocytes and BMP15 transcript increased significantly (P<0.05) between 6 h and 12 h and decreased again between 12 h and 24. In cumulus cells, GDF9 remained stable during IVM upto 12 h of maturation and decreased significantly between 12 h and 24 h of maturation. Conversely, significant reduction of BMP15 was observed between 0 h and 6 h, stayed stable upto 12 h and became undetectable at 24 h of maturation. In conclusion, these two genes were differentially expressed during the period of oocyte maturation process and notably, BMP15 expression pattern is associated specifically with the period of cumulus cell expansion.
