Effect of endothelial progenitor cells on the proliferation of co-cultured vascular smooth muscle cells
10.3969/j.issn.1672-7347.2010.01.008
- VernacularTitle:内皮祖细胞对血管平滑肌细胞增殖的影响
- Author:
Li FANG
;
Meifang CHEN
;
Guolong YU
;
Zhilin XIAO
;
Xiaobin CHEN
;
Xiumei XIE
- Publication Type:Journal Article
- Keywords:
endothelial progenitor cell;
vascular smooth muscle cell;
proliferation;
cell cycle
- From:
Journal of Central South University(Medical Sciences)
2010;35(1):52-62
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of endothelial progenitor cells (EPCs) on the proliferation of co-cultured rat vascular smooth muscle cells(VSMCs). Methods Mononuclear cells were isolated from fresh cord blood by 6% hydroxyethyl starch (HES) and density gradient centrifugation. Isolated mononuclear cells were cultured in EGM-2 medium supplemented with 20% fetal bovine serum (FBS), VEGF,bFGF and other growth factors. Biological features of EPCs were observed at different time points, and EPCs were identified by morphology, fluorescence double-staining and flow cytometry. Indirect immunofluorescence was performed to analyze the expression of α-SM-actin, calponin of VSMCs special antigen. Co-culture system of EPCs and VSMCs was established by transwell permeable support. FBS (20%) was used to stimulate the proliferation of VSMCs. In a VSMCs/EPCs co-culture system, the DNA synthesis ability, total protein level and cell cycle of VSMCs were determined by BrdU marking method,protein quantitation and flow cytometry after co-culture for 6, 12, 24,48 and 72 h. Results After co-culture for 12, 24, 48, and 72 h, the DNA synthesis ability and total protein level of VSMCs significantly decreased compared with the control group(P＜0.05). Flow cytometry showed that the percentage of S phase of VSMCs in VSMCs/EPCs co-cultured group significantly decreased and the percentage of G_1 phase increased markedly compared with the control group(P＜0.05). The maximal inhibitory effect was observed at 48 h. Conclusion Early EPCs could inhibit the proliferation of VSMCs.
