Cell-cell contact induces mesenchymal stem cells differentiation to smooth muscle cells
- VernacularTitle:细胞间接触诱导骨髓间充质干细胞向平滑肌细胞分化的实验研究
- Author:
Xiaofang LIU
;
Yanzhou WANG
;
Huicheng XU
;
Zhiqing LIANG
- Publication Type:Journal Article
- Keywords:
cell-cell contact;
microenvironment;
mesenchymal stem cells;
smooth muscle cells
- From:Journal of Third Military Medical University
1988;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a co-culture system between mesenchymal stem cells (MSCs) and smooth muscle cells (SMCs) in vitro, and study the influence of SMCs on the differentiation of bone marrow MSCs (BMSCs) into SMCs in the co-culture system. Methods Density gradient centrifugation was used to separate and culture BMSCs, and collagenase digestion was employed to separate and culture bladder smooth muscle cells(BSMCs). Then the 2 types of cells were identified respectively. Then the cells were divided into 4 groups, BMSCs culture group (positive control), BSMCs cells culture group (negative control), BMSCs and BSMCs co-culture groups with contact or non-contact. The expression of ?-smooth muscle actin (?-SMA) and calponin were detected by immunofluorescence staining. Western blot analysis was employed to determine protein expression of calponin. Results In contact co-culture group, the positive rate of CD90 abd CD44 were 99.78% and 60.27% respectively in BMSCs, while only 0.21% for CD45. Western blotting showed that BMSCs expressed little calponin, but those contacting with BSMCs had an increasingly stronger expression of calponin with the elapse of contact culture time, while there was no significant change in its expression in BMSCs with contact. Immunofluorescence staining indicated that part of BMSCs were calponin positive at the 8th day after culture, but no positive cell was observed in non-contact culture group and BMSCs culture group. Conclusion The contact between BSMCs and BMSCs can promote the differentiation of BMSCs into SMCs.