The feeder layer of human embryonic fibroblasts supports the growth of human spermatogonial stem cells.
- Author:
Yu-Bin WANG
1
;
Bin CHEN
;
Ying-Chao WANG
;
Zhi-Ling ZHANG
;
Hong-Xiang WANG
;
Yong-Ning LU
;
Zu-Qiong XIANG
;
Kai HU
;
Yi-Ke YANG
;
Yin-Fa HAN
;
Zheng WANG
;
Yi-Xin WANG
;
Yi-Ran HUANG
Author Information
- Publication Type:Journal Article
- MeSH: Cell Culture Techniques; methods; Cell Differentiation; Cells, Cultured; Embryo, Mammalian; cytology; Fibroblasts; cytology; Humans; Male; Spermatogonia; cytology; Stem Cells; cytology
- From: National Journal of Andrology 2008;14(12):1063-1068
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the methods and conditions for the isolation, purification and culture of human spermatogonial stem cells (SSCs) on the feeder layer cells of human embryonic fibroblasts (hEFs).
METHODSSSCs isolated and purified from normal human fetal testicular tissues by sequential two-step enzyme digestion and Percoll uncontinuous density gradient centrifugation were cultured on the feeder layer cells of hEFs isolated from 5-9 weeks old human embryos. The surface markers SSEA-1 and OCT4 of the SSCs were detected by immunohistochemistry; the alkaline phosphatase (AKP) activity of the SSC clones measured; and the expressions of the SSC-related genes determined by RT-PCR.
RESULTSSSCs survived, proliferated and formed colonies on the feeder layers, and the colonies were highly positive for SSEA-1 and OCT4, with strong AKP activity and high expressions of the SSC-related genes.
CONCLUSIONThe feeder layer of hEFs supports the growth of human spermatogonial stem cells.
