[Experimental study of differentiation of canine bone marrow mesenchymal stem cell into fibroblasts in vitro].
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.
Wei
Xiang-yang
XY
Department of Cardiovascular Surgery, Xijing Hospital, the Fourth Military Medical University, Xi'an 710032, China.
Liu
Wei-yong
WY
Sun
Guo-cheng
GC
Ouyang
Hui
H
Gu
Chun-hu
CH
Liu
Xing-guang
XG
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