Isolation,Culture and Multiple Differentiation Induction of Nanyang Bovine Bone Marrow Mesenchymal Stem Cells

The study aimed to build the method of Nanyang bovine bone marrow-derived mesenchymal stem cells (BMSCs) separation culture in vitro, and study its bionomics and the ability of multiple differentiation induction on this basis. Using bone marrow puncture to take rib bone marrow of 3 months calf, isolation and culture BMSCs, subcultured and determined its curve of growth, detected Oct4, Nanog, Sox2 genes expression by RT-PCR technique, then took P3 BMSCs into neural and fat cells to carry out induction differentiation, and took the use of histology staining technique and RT-PCR technique to detect. The results of the BMSCs were uniform sindleshaped in appearance; RT-PCR could dectect the expression of stem cell factor Oct4, Nanog, Sox2. The results showed that an "S" shape growth curve of cell at different generations times, it entered exponential growth stage at the 3rd day generally, and entered plateau after then the 7th day. Into nerve after induction, stained with toluidine blue was apparent throughout the structure of Nissl substance, ENO2 and GFAP genes showed a positive expression. After adipogenic differentiation of ASCs was assessed by oil Red O staining which showed a large number of lipiddroplet, RT-PCR dectected Leptin and PPAR genes showing a positive expression. The tests showed that had successfully isolated from Nanyang bovine BMSCs and it had the potential of multidirectional differentiation.