[Experimental study on differentiation of adult marrow mesenchymal stem cells into vascular endothelial cells in vitro].
OBJECTIVE
To study the differenation of adult marrow mesenchymal stem cells (MSCs) into vascular endothelial cells in vitro and to explore inducing conditions.


METHODS
MSCs were isolated from adult marrow mononuclear cells by attaching growth. MSCs were divided into 4 groups to induce: the cells seeded at a density of 5 x 10(3)/cm(2) in 2% and 15% FCS L-DMEM respectively (group 1 and group 2), at a density of 5 x 10(4)/cm2 in 2% and 15% FCS L-DMEM respectively (group 3 and group 4); vascular endothelial growth factor (VEGF) supplemented with Bovine pituitary extract was used to induce the cell differentiation. The differentiated cells were identified by measuring surface marks (CD34, VEGFR-2, CD31 and vWF ) on the 14th day and 21st day and performed angiogenesis in vitro on the 21st day. The cell proliferation index (PI) of different inducing conditions were measured.


RESULTS
After induced in VEGF supplemented with Bovine pituitary extract, the cells of group 3 expressed the surface marks CD34, VEGFR-2, CD31 and vWF on the 14th day, the positive rates were 8.5%, 12.0%, 40.0% and 30.0% respectively, and on the 21st day the positive rates of CD34 and VEGFR-2 increased to 15.5% and 20.0%, while the other groups did not express these marks; the induced cells of group 3 showed low proliferating state (PI was 10.4%) and formed capillary-like structure in semisolid medium.


CONCLUSIONS
Adult MSCs can differentiate into vascular endothelial cells after induced by VEGF and Bovine pituitary extract at high cell densities and low proliferating conditions, suggesting that adult MSCs will be ideal seed cells for the therapeutic neovascularization and tissue engineering.
Yan
Kaili
K
Department of Central Experimental Laboratory, Anhui Provincial Hospital, Hefei Anhui, 230000, P. R. China.
Wang
Jian
J
Li
Qing
Q
D000328:Adult; D018952:Antigens, CD34 / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism
D018952:Antigens, CD34; D040301:Vascular Endothelial Growth Factor Receptor-2