Optimization of the culture system for the mesenchymal stem cells from the placenta chorion in vitro and the preliminary analysis of its biological characterization / 中国免疫学杂志

ABSTRACT

Objective:To optimize the culture system for the mesenchymal stem cells from the Placenta Chorion (hCMSC) in vitro by adding the selected cytokines to amplify the hCMSC effectively,with furthermore some preliminary analysis of the hCMSC performed.Methods:Immunohistochemistry was applied to show the localization of the mesenchymal stem cells within the placenta chorion,then the corresponding part of chorion was digested by collagenase Ⅳ to get the cell suspension for primary culture and subculture;Flow Cytometry was used to detect the surface marker of hCMSC,whose proliferation was detected by direct cell counting and the MTT method;then Butylated Hydroxyanisole (BHA),and DMSO was used combinedly as the inducer in the differentiation of the hCMSC to the neuron like cells,which was then detected by Immunofluorescence staining for the expression of NSE;finally RT-PCR was used to detect the expression of flt3.Results:The result of immunohistochemistry showed CD105~-,CD90~- positive hCMSC was much easier to be found around the blood vessels within the villous of chorion,which then could be obtained by mechanical dissociation and collagenase digestion.bFGF was good at amplifying the hCMSC among all selected cytokines,but FL was showed even obviously effective on the proliferation of the hCMSC.The propagating cells were found to highly express CD73,CD90,CD105;and negative for CD14,CD34,CD45 and HLA-DR.We also found that the hCMSC happened to express PDL-1 the negative co-stimulatory molecules,while their potential to different into the neuron like cells was still well maintained.Conclusion:There is low immunogenicity mesenchymal stem cells within the placenta chorion,both bFGF,FL are shown good effects on the proliferation of the hCMSC,with maintaining their differentiation potential as well.