Study On Isolation,Culture And Multiple Differ,Entiation Potential Of Human Umbilical Cord Mesenchymal Stem Cells

Keratinocyte is the main cell type in epidermal skin tissues.And it is the most important cell type in the cell therapy field of skin injury as well.Mesenchymal stem cell is one of the ideal seed cell types in cell therapy.It has very low immunogenicity and multi-directional differentiation ability.In this study,the plenty numbers of human.umbilical cord mesenchymal stem cells were effieiently collected from the Wharton’s jelly tissue pieces,isolated from unmbilical cord,which clung on the bottom of dish for the second time,which was a meliorated method upon usual ways.They finally differentiated into various kinds of cells by cultured in special medium.The main results were as follow:1.Human umbilical cord mesenchymal stem cell line was established.The cells grew in a circinate regulation with long-fusiform shapes.The 3rd passage hUCMSCs expressed Sox2 gene,which was one of the stem cell transcription factor,and also expressed CD29,CD90 and CD105 gene,but absented of CD34,CD45 and HLA-DR.The growth curve showed an "S" shape,and the population doubling time is 24.7 hours.These cells showed a favourable growing condition with normal male karyograms(46,XY)and heredity was stable.2.The multi-directional differentiation systems of hUCMSCs were established.hUCMSCs were inducted to differentiated into ectoderm(neuron and keratinocyte)and mesoderm(osteoblast)cell types.All the staining results were positive.3.Keratinocyte-differentiation medium were created.After cultured in this medium for a period of time,hUCMSCs were differentiated into keratinocytes.They expressed cytokeratin,observed in immunological staining results.And the expression of the special gene CK19 was increased day by day in the molecular level testing.In summary,this study found a new way to obtain hUCMSCs,’and established UCMSC line which has the multi-directional differentiation potential.A keratinocyte differentiation medium was also found,This research provided experimental basis to clinical application of mesenchymal stem cells in cell therapy especially the large area skin injury.