title: [Isolation, culture, and identification of human spermatogonial stem cells].
abstract: OBJECTIVE
  To isolate, identify and culture human spermatogonial stem cells (SSC) and then obtain purified and enriched human SSCs for research and application.
  
  
  METHODS
  We detected the expression of CD90 in the human testis using the immunofluorescence technique and isolated human testicular spermatogenic cells by two-step enzymatic digestion, followed by differential plating and magnetic-activated cell sorting (MACS) with CD90 as an SSC marker. Then we identified the isolated CD90-positive spermatogenic cells by RT-PCR and immunocytochemistry, and meanwhile cocultured them with Sertoli cells in SG medium in vitro.
  
  
  RESULTS
  The isolated CD90-positive cells showed a relatively homogeneous characteristic in size and morphology and expressed the genes specific for human SSCs, with high expressions (90.5%) of GFRA1, GPR125, and UCHL1. After coculture with Sertoli cells in the SG medium for 2 weeks, the isolated CD90-positive cells maintained a good activity.
  
  
  CONCLUSIONS
  CD90 can be regarded as a speci- fic marker for human SSCs and used to obtain highly enriched human SSCs by differential plating and MACS. Furthermore, the isolated human SSCs can be cultured in SG medium in vitro.
authors.forename: Jun-long
authors.initials: JL
authors.lastname: Wang
authors.forename: Shi
authors.initials: S
authors.lastname: Yang
authors.forename: Ru-hui
authors.initials: RH
authors.lastname: Tian
authors.forename: Zi-jue
authors.initials: ZJ
authors.lastname: Zhu
authors.forename: Ying
authors.initials: Y
authors.lastname: Guo
authors.forename: Qing-qing
authors.initials: QQ
authors.lastname: Yuan
authors.forename: Zu-ping
authors.initials: ZP
authors.lastname: He
authors.forename: Zheng
authors.initials: Z
authors.lastname: Li
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology*; D015415:Biomarkers / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D048430:Cell Shape; D048429:Cell Size; D018920:Coculture Techniques; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D012708:Sertoli Cells; D013093:Spermatogonia / Q000166:cytology*; D013737:Testis / Q000378:metabolism; D018800:Thy-1 Antigens / Q000302:isolation & purification / Q000378:metabolism*; D043222:Ubiquitin Thiolesterase / Q000378:metabolism
chemical_list: C548160:ADGRA3 protein, human; D015415:Biomarkers; C494987:GFRA1 protein, human; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; D043562:Receptors, G-Protein-Coupled; D018800:Thy-1 Antigens; C482860:UCHL1 protein, human; D043222:Ubiquitin Thiolesterase