Research On Isolation, Culture And Differentiation Potential Of Chick Embryo Dermis-derived Stem/Progenitor Cells

In the 90th of last century,the stem cells were studied by scientists around the world.The Scientists paid more and more attention to stem cells about their ability of proliferation and differentiation.Nowadays the research on stem cells not only included isolation and culture,but also included the differentiation potency and Clinical application.In this study,the culture system of Dermis-derived Mesenchymal Stem/ progenitor Cells (DMS/PCs) were established and the differentiation potency of DMS/PCs was explored, The results were in the below:1. The skin tissue was isolated from 16 days old Beijing fatty chick embryo,and the dermal layer was isolated from epidermal layer by digested with 0.25% DispaseⅡfor about 1.5-2h at 37℃. The DMS/PCs were gained by digested with 0.25% trypsin solution for about 15 min.The suitable culture condition for DMS/PCs were L-DMEM medium,20ng/ml EGF,20ng/ml b EGF,2mM L-glutamine and 1mM sodium pyruvate.2.Beijing fatty chicken DMS/PCs self-renewal were very strong in vitro and exhibited a fibroblast-like morphplogy. Primary culture cells were mixed with other cells, for example blood cells and fibroblasts, DMS/PCs can be purified after 2-3 passages. The duplication time of Beijing fatty chicken DMS/PCs was 72h.The cells can be passeged to 16 generations. Cell morphology and proliferation were relatively stable within 13 generations, starting from the 14th generation, the cells aged significantly and bubbled, and the cells which sub-cultured to the 16th passage were not proliferous and plenty of cells were death.3. Cultured cells were detected of markers by immunofluorescence staining and RT-PCR method.The results showed the cells expressed CD29, CD44, CD71, CD73 which express in most mesenchymal stem cells.4. Under the appropriate effect of inducing factors, Beijing fatty chicken DMS/PCs could be induced to differentiate into osteoblasts, adipocytes and even neurocytes.In summation, separation and culture of DMS/PCs had active proliferative ability and multiple differentiation potentials, at the same time, DMS/PCs were easy to acquire and cultured in vitro,because of these advantages, DMS/PCs could be used not only to be preservation of endangered species and clinical application,but also be useful to Genetic Improvement and Optimization of Beijing fatty chicken.