Endometrial stromal stem cells:in vitro isolation, culture and biological features

Bi Yu-hu; Teng Jun-ru

doi:10.3969/j.issn.2095-4344.2017.05.018

Abstract：BACKGROUND:Understanding and studying endometrial cel s from the perspective of stem cel s can provide a new therapeutic approach and research entry point for the clinical treatment of endometrial diseases. OBJECTIVE:To compare the biological features of endometrial stromal stem cel s isolated and cultured using different methods. METHODS:Three commonly used cel separation methods, including trypsin, col agenase I and their combination, were used to isolate and culture endometrial stromal stem cel s from the uterus after removal. Afterwards, passage 2 cel s cultured for 5 days were subjected to trypan blue staining and living cel counting. Immunohistochemical staining for CD146 and CD90 and RT-PCR were used to identify harvested endometrial stromal stem cel s. Logarithmical y growing cel s were cultured to draw cel growth curves. RESULTS AND CONCLUSION:Harvested endometrial stromal stem cel s presented with spindle, adherent cel growth, nonpolar arrangement, and fibroblast morphology. The DNA of endometrial stem cel marker CD146 andCD90 highly expressed in 500 bp and 150 bp respectively, but did not express in the mesometrium. S-shaped growth curve of cultured cel s was found, and there was no difference in the cel cycle of cel s cultured using three different methods. Among the three methods, col agenase I method could harvest the highest number of endometrial stromal stem cel s that grew fastest. These findings indicate that endometrial stromal stem cel s can be successful y isolated from the removed uterine tissues, and col agenase I method is an efficient, practical and stable method for in vitro culture of endometrial stromal stem cel s.