[Isolation, culture and differentiation into endothelial-like cells from rat bone marrow mesenchymal stem cells in vitro].
OBJECTIVE
To study isolation and culture from SD rat bone marrow mesenchymal stem cells (rBMMSCs) and differentiate into endothelial-like cells (ELCs) with VEGF and bFGF.


METHODS
In vitro rBMMSCs were cultured with the method of percoll (1.073 g/ml) density centrifugation and adherence to plastic dishes. Then they were seeded in LG-DMEM supplemented with 10% fetal bovine serum. The relative biologic characteristics of rBMMSCs including cell morphology, phenotype, growth curve, cell cycle and ultrastructure were studied by inverted microscope, immunocytochemistry, flow cytometry, MTT method, transmission electron microscopy(TEM). The induced cells were identified by immunocytochemistry with CD31, CD34, CD144(VE-cadherin), FITC-UEA-1 and had ability in Dil-ac-LDL uptake and were observed under inverted microscope.


RESULTS
The morphology of rBMMSCs was spindle and has whirlpool. Growth curve of P4 showed that there was difference of latency, activity and flat periods. TEM showed that there were two of rMSCs , the small of both were infantile cells. GO/G1 phase of cell cycle was 95.67% and was suggested that most of cells were not in period of proliferation. The part differentiated cells demonstrated the characters of endothelial-like cells under inverted microscope and showed the expression of CD31, CD144, CD34, and possed the functions of endothelial-like cells staining double-positive for Dil-Ac-LDL and FTIC-UEA-1.


CONCLUSIONS
1.073 g/ml percoll density centrifugation and cultured adherence is an effective approach to obtain rBMMSCs. In vitro, the cells have potential to differentiate into endothelial-like cells
Zhao
Ying
Y
College of Bioengineering, Chongqing University, Chongqing 400044, China.
Cai
Shao-xi
SX
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation / Q000502:physiology*; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology*; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051381:Rats; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology
D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2