Differentiation Potential and Culture Requirements of Mesenchymal Stem Cells from Ovine Bone Marrow for Tissue Regeneration Applications

Objectives- To isolate, culture-expand and differentiate mesenchymal stem cells from ovine bone marrow and determine their culture requirements for high expansion rate. Design- Experimental study. Animals- Five Shal sheep. Procedures- In this study, ovine marrow cells were plated and culture expanded through 3 successive subcultures. The resultant cells were then plated at differentiating conditions into bone, cartilage and adipose cell lineages in order to verify their MSC nature. Furthermore, we determined the culture requirements of the cells in terms of FBS (fetal bovine serum) concentration and initiating cell seeding density. In this study, also population doubling time (PDT) was determined for the isolated cells. Results- According to our observations, MSCs from ovine marrow appeared to be fibroblastic in appearance. They were easily able to differentiate into bone, cartilage and adipose cell lineages as it was evident in RT-PCR analysis of specific gene expression and specific staining of differentiated cells. According to our findings, the cells indicated extensive proliferation when being cultivated at 100 cells/cm2 in a medium with 15% FBS. Ovine MSCs possessed a relatively short population doubling time (24.94±2.67 hours). Conclusions and Clinical Relevance- Fibroblastic cells from ovine bone marrow are able to undergo extensive proliferation and capable of differencing into three skeletal lineages, hence they are MSCs that are appropriate for cell therapy experimentation