Authors : Deying Zhang, Guanghui Wei, Dawei He, Jing Zhu, Xing Liu and Tao Lin
Abstract: Spermatogonial Stem Cells (SSCs) are the unique stem cells in the body that can transmit genetic information to the offspring, which are recently demonstrated to be with more perspectives in cell engineering. In this study, an easy and feasible method of rat SSCs in vitro culture system is established which can facilitate the study on SSCs and the biological characteristics of rat SSCs cultured in vitro were observed. Sertoli cells and SSCs were separated by two-step enzyme digestion from 0-1 and 6-7 days old Sprague-Dawley rats, respectively. SSCs were inoculated on the Sertoli cells on the 5th day when the feeder layer formed. Morphology of coculture cells was observed and expression of SSCs markers Vasa, SMAD1, Stella, Dazl and GCNF, HSP90α, Oct-4, integrin-β1 were detected by RT-PCR and Western blot respectively. SSCs began to proliferate and differentiate 24 h after being cultured on the Sertoli cells layer and there were a few A-paired (Apr) spermatogonia cell colonies. With time prolongation, the number of Apr cell colonies decreased gradually, meanwhile the number of aligned (Aal) cell colonies increased, then Aal cell colonies retained stable quantity after 120 h in culture, it could retain (54.1±6.2 days) under the condition the culture medium being changed every 4-5 days. We concluded that rat SSCs can proliferate colonially on the feeder layer of Sertoli cells and retain stable morphous and quantity. The SSCs culture system fed by Sertoli cells layer can serve as a tool to faciliate SSCs and SSCs-Sertoli cells interaction research.
Deying Zhang, Guanghui Wei, Dawei He, Jing Zhu, Xing Liu and Tao Lin, 2010. Biological Characteristics of Rat Spermatogonial Stem Cells Cultured on Sertoli Cell Feeder Layer in vitro. Journal of Animal and Veterinary Advances, 9: 1765-1770.