The Establishment And Chondrocyte-differentiation Of Equine Bone Marrow Mesenchymal Stem Cells

In this study, we established equine BMSCs, identified its biological characteristics and stem cell characteristics (CD34、 CD44、CD45、CD90、CD105、 SOX2、Nanog), then we induced BMSCs differentiate into chondrocytes in some special culture. After alcian blue staining and RT-PCR gene identification with Collagen type Ⅱ, we confirmed that the differentiated cells performanced chondrocyte-specific morphology and expressed specific genes. The results are as follows:1. We established equine bone marrow mesenchymal stem cell lines. BMSCs are shuttle-shaped, just like the fibrocytes. The P3 BMSCs growth curve showed a "S" type, and the doubling time is about 31 hours.2. After gene identification by PCR, the P3 BMSCs express stem cell special factors, such as SOX2, NANOG, CD14, CD90, CD105, but absent of CD34 CD45.3. We cultured BMSCs in chondrocyte-differentiated medium and ordinary medium respectively for about 21 days, after staining two group of cells with alcian blue, indicated the presence of glycoprotein in differentiated cells and the morphology of chondtocytes is very obvious, but control group cells were not been stained with alcian blue and were still showed a shuttle shape. After qRT-PCR,we found that the differentiated cells express Collagen type Ⅱ, but the control cells do not.To sum up, we have established equine bone marrow mesenchymal stem cells successfully, and induced into chondrocytes. Our research results provide both the theory foundation and technologies for cell therapies about equine athletic injuries, as well the human clinical medicine.