title: [The feeder layer of human embryonic fibroblasts supports the growth of human spermatogonial stem cells].
abstract: OBJECTIVE
  To 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).
  
  
  METHODS
  SSCs 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.
  
  
  RESULTS
  SSCs 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.
  
  
  CONCLUSIONS
  The feeder layer of hEFs supports the growth of human spermatogonial stem cells.
authors.affiliation: Department of Urology, Shanghai Institute of Andrology, Shanghai, China. wangyb1980@163.com
authors.forename: Yu-Bin
authors.initials: YB
authors.lastname: Wang
authors.forename: Bin
authors.initials: B
authors.lastname: Chen
authors.forename: Ying-Chao
authors.initials: YC
authors.lastname: Wang
authors.forename: Zhi-Ling
authors.initials: ZL
authors.lastname: Zhang
authors.forename: Hong-Xiang
authors.initials: HX
authors.lastname: Wang
authors.forename: Yong-Ning
authors.initials: YN
authors.lastname: Lu
authors.forename: Zu-Qiong
authors.initials: ZQ
authors.lastname: Xiang
authors.forename: Kai
authors.initials: K
authors.lastname: Hu
authors.forename: Yi-Ke
authors.initials: YK
authors.lastname: Yang
authors.forename: Yin-Fa
authors.initials: YF
authors.lastname: Han
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.forename: Yi-Xin
authors.initials: YX
authors.lastname: Wang
authors.forename: Yi-Ran
authors.initials: YR
authors.lastname: Huang
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D008297:Male; D013093:Spermatogonia / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*