title: [Isolation and culture of fetal bovine intestine-derived epithelial stem cells and the differentiation into hepatocyte-like cells].
abstract: OBJECTIVE
  To establish the culture system of fetal bovine intestinal epithelial stem cells (IESCs) in vitro, identify specific markers of the cell lines and analyze the differentiation potential into hepatocyte-like cells.
  
  
  METHODS
  IESCs were isolated from the 3- to 5-month fetal bovine intestine by the digestion of collagenase I, and cultured in the DMEM/F12 medium. The cell morphology was observed, and the proliferation ability and multiple differentiation potential were demonstrated by subculturing and its growth curve. The mRNA expressions of the surface markers Bmi1, Hes1, Lgr5 and cytokeratin 19 (CK19) were determined by reverse transcription PCR (RT-PCR), and the protein levels of Bmi1, LGR5 and CK19 were detected by immunofluorescence cytochemistry. Under the induction of fibroblast growth factor 4 (FGF-4) and hepatocyte growth factor (HGF), the cell differentiation into hepatocyte-like cells was assayed by the glycogen staining and RT-PCR.
  
  
  RESULTS
  IESCs cultured in vitro expressed Bmi1, Hes1, Lgr5 and CK19 mRNAs, and CK19, Bmi1 and LGR5 proteins. The differentiated cells were positively stained by glycogen, and RT-PCR showed that the cells expressed α-fetoprotein (AFP) and albumin (ALB) mRNAs.
  
  
  CONCLUSIONS
  The culture system of IESCs in vitro is successfully established, and the cells are differentiated into hepatocyte-like cells.
authors.affiliation: College of Basic Medicine, Jiamusi University, Jiamusi 154007; China Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Sun
authors.affiliation: College of Basic Medicine, Jiamusi University, Jiamusi 154007, China.
authors.forename: Lianshun
authors.initials: L
authors.lastname: Cai
authors.affiliation: China Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Weijun
authors.initials: W
authors.lastname: Guan
mesh_terms: D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D051519:Fibroblast Growth Factor 4 / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000166:cytology* / Q000196:embryology; D053539:Keratin-19 / Q000235:genetics / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D051519:Fibroblast Growth Factor 4; D053539:Keratin-19; D043562:Receptors, G-Protein-Coupled