title: [Construction of directional differentiation model from mouse embryonic stem cells to Leydig-like cells in vitro].
abstract: OBJECTIVE
  To construct a directional differentiation model from mouse embryonic stem cells into leydig-like cells in vitro.
  
  
  METHODS
  Mouse ES-D3 cells were transfected with plasmid containing steroidogenic factor 1 (SF-1) gene, then treated with RA and 8Br-cAMP, while the cells transfected with empty plasmid were used as the negative controls. The morphology of leydig-like cells differentiated from ES-D3 cells was observed with light microscopy. The expression levels of StAR, P450scc and 3β-HSD were detected by RT-PCR, Western Blot and fluorescence microscopy analysis in leydig-like cells derived from the ES cells.
  
  
  RESULTS
  ES-D3 cells were transfected with plasmid containing SF-1 gene successfully, and SF-1 was expressed 24 h after transfection. The SF-1-transfected ES-D3 cells were induced by RA and 8Br-cAMP to differentiate into leydig-like cells. The differentiated cells showed spindle shape with tentacles, which expressed the specific protein marker for leydig cells 3β-HSD1 and P450scc. Meanwhile, in these leydig-like cells, the expression of StAR increased compared with control group. 3β-HSD1, P450scc and StAR were not detected in negative control group.
  
  
  CONCLUSIONS
  When the ES-D3 cells are transfected with SF-1 plasmid and then treated with RA and 8Br-cAMP, the cells are able to differentiate into leydig-like cells, indicating that the model of directional differentiation of ES cells into leydig-like cells has been constructed successfully.
authors.affiliation: Institute of Pharmacology, Toxicology and Biochemical Pharmaceutics, Zhejiang University, Hangzhou 310058, China.
authors.forename: Ying-ying
authors.initials: YY
authors.lastname: Zhang
authors.forename: Ya-dong
authors.initials: YD
authors.lastname: Huang
authors.forename: Ren-shan
authors.initials: RS
authors.lastname: Ge
authors.forename: Dan-yan
authors.initials: DY
authors.lastname: Zhu
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D007985:Leydig Cells / Q000166:cytology* / Q000378:metabolism; D008297:Male; D051379:Mice; D054339:Steroidogenic Factor 1 / Q000235:genetics*; D014162:Transfection
chemical_list: D054339:Steroidogenic Factor 1