Biological Characteristics of Mesenchymal Stem Cells Cultured in Conditions for Spermatogonia in Vitro / 实用儿科临床杂志

ABSTRACT

Objective To investigate the possibility of inducing mesenchymal stem cells(MSCs)from human umbilical cord Wharton's Jelly to differentiate into spermatogonia.Methods To isolate,culture and purify MSCs with adherent method,the growth and proliferation of human umbilical cord-derived MSCs were observed,and their immunophenotypes were determined by flow cytometry;MSCs of the third generation were divided into 2 groups to be induced and cultured,MSCs of the control group were cultured in basal medium,while those of the experimental group with conditional medium.The morphologic and ultrastructure changes of control group and experimental group cells were compared with phase contrast microscopy,electron microscopy(EM)and transmission electron microscope(TEM)respectively ;the spermatogonial cells differentiated were then evaluated by immunohistochemistry stained for CD117and CD49f ;the method of Western-blot was used to test if the cells induced could express CD49f.Results A population of MSCs were isolated from human umbilical Wharton's Jelly;they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling;After induction,the shape of MSCs changed greatly from the fibroblast to the round,even familiar to the tadpole;expressed the known molecular markers of spermatogonial cells,such as CD49f,CD117.Conclusion The induced MSCs not only undergo spfermatogonial-cell like morphologic changes,ultramicrostructure mature with increasing cell organs,but also express the spermatogonial cell markers,which show that human umbilical cord derived MSCs are capable of differentiating into spermatogonial cell.