Preliminary Research On Isolation And Multilineage Potential Of Mesenchymal Stem Cells Derived From Canine Umbilical Cord And Giant Panda Bone Marrow

Stem cells are known as a population of self-renewal and differentiation potential cells. According to its differentiation stage,the stem cells can be divided into embryonic stem cells and adult stem cells which contained the mesenchymal stem cells. The present study, mesenchymal stem cells are mostly derived from human or animal tissue and have good research and application prospects because of its availability, can be amplified and differentiation ability. But the current research on animals in this area is still relatively small. In this study MSCs were successfully isolated from the umbilical cords of Canine and the bone marrow of Giant panda.The research characteristics of these cells, e.g., morphologic appearance, growth curve, differentiation potential and gene expression were investigated. The results are as follows:A:Isolation,culture, differentiation potential of mesenchymal stem cells derived from the umbilical cords of CanineFibroblast-like cells both can be isolated no matter using tissue culture or enzyme digestion.The cells from tissue culture their vitality and proliferation rate are better than the cells from enzyme digestion,they could stably growth to the19th passages,but the enzyme digested cells dead in the second passages.The tissue culture cells grew in parallel or swirling.The cell growth curve was S-shaped and cell population doubling time was about57h. The assays in vitro demonstrated the cells exhibited multi-potential differen-tiation into osteogenic and adipogenic cells.We could find red calcium nodules by alizarin red staining after osteogenic differentiation and red oil drops by oil red O staining after adipogenic differentiation. All the assays demonstrated the cells derived from the umbilical cords of Canine were mesenchymal stem cells.B:Isolation,culture, differentiation potential of mesenchymal stem cells derived from the bone marrow of Giant pandaThe fibroblast-like cells were successfully derived from the bone marrow of Giant panda by using whole bone marrow culture method. The cells grew in parallel or swirling and could stably growth to the10th passages. The cell growth curve was S-shaped and cell population doubling time was about60h. The assays in vitro demonstrated the cells exhibited multi-potential differentiation into osteogenic and adipogenic cells. We could find red calcium nodules by alizarin red staining and blue cells by alkaline phosphatase staining after osteogenic differentiation and red oil drops by oil red O staining after adipogenic differentiation. RT-PCR analysis showed the cells expressed CD73、SOX-2, but there was no expression of CD105、CD90after immunohistochemical analysis. Based on the above studies, We speculated that the cells derived from the bone marrow of Giant panda were mesenchymal stem cells.