title: [Experimental study of differentiation of canine bone marrow mesenchymal stem cell into fibroblasts in vitro].
abstract: OBJECTIVE
  To explore the way of stably inducing canine bone marrow mesenchymal stem cells (BMSCs) to differentiate into fibroblasts and myofibroblasts in vitro, and provide seed cells for fabricating tissue engineering heart valves (TEHV).
  
  
  METHODS
  Adult canine BMSCs were separated by a gradient centrifugation on Percoll (density 1.073 g/ml), then the cells were incubated in low-glucose Dulbecco Eagle's minimum essential medium (LG-DMEM) with 10% bovine calf serum. Cell phenotype were identified by immunohistochemistry staining. The second and third generation of BMSCs were committedly induced by conditioning culture medium, which were detected by immunohistochemistry staining. The induced-BMSCs were freezed, preserved and resuscitated after 7 d to observe the cell growth, proliferation and function.
  
  
  RESULTS
  BMSCs deriving from the bone marrow mononuclear cells separated by a Percoll gradient were positive expression of alpha-smooth muscle antibody, vimentin and negative expression of CD34, laminin. About (50 +/- 3)% induced-BMSCs were positive expression of laminin. Approximately (85 +/- 3)% freezed induced-BMSCs could be resuscitated. And the growth, proliferation and function were well.
  
  
  CONCLUSIONS
  BMSCs could be committedly induced to differentiate into fibroblasts and myofibroblasts in vitro. It is suitable to be the seed cells.
authors.affiliation: Department of Cardiovascular Surgery, Xijing Hospital, the Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Xiang-yang
authors.initials: XY
authors.lastname: Wei
authors.forename: Wei-yong
authors.initials: WY
authors.lastname: Liu
authors.forename: Guo-cheng
authors.initials: GC
authors.lastname: Sun
authors.forename: Hui
authors.initials: H
authors.lastname: Ouyang
authors.forename: Chun-hu
authors.initials: CH
authors.lastname: Gu
authors.forename: Xing-guang
authors.initials: XG
authors.lastname: Liu
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D004285:Dogs; D005347:Fibroblasts / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D009000:Monocytes / Q000166:cytology*; D032446:Myoblasts / Q000166:cytology