目的:研究骨髓基质干细胞来源的血管内皮细胞表达成熟血管内皮细胞特异性基因,探讨为组织工程血管化提供种子细胞的可行性.方法:从SD大鼠后肢获取骨髓基质干细胞,进行体外扩增培养,取生长良好的第3代细胞体外诱导培养14 d.细胞分诱导组(含内皮诱导因子VEGF和bFGF的完全血管内皮细胞条件培养基)和对照组(含M199普通培养基).MTT法检测两组细胞增殖活性.免疫细胞化学法检测CD31,CD34蛋白的表达;RT-PCR检测细胞血管生成素1(Ang1)、血管生成素2(angiogenin2,Ang2)mRNA的表达.结果:MIT法检测发现,诱导组与对照组在1,2,3,4,5 d 5个时间点内,其细胞增殖活性无显著性差异(P_(1d)=0.855,P_(2d)=0.053,P_(3d)=0.249, P_(4d)=0.059,P_(5d)=0.174).免疫细胞化学检测结果CD34,CD31呈阳性表达.RT-PCR结果显示,诱导的血管内皮细胞表达成熟内皮细胞特异性基因Ang1,Ang2.结论:骨髓基质干细胞在一定诱导条件下可以表达血管内皮细胞特异性标志,可作为组织工程血管化理想的种子细胞.
Gene expression of angiogenin1 and an giogenin2 of endothelial cells derived from rat bone marrow mesenchymal stem cells
AIM: To investigate the specific genes of vascular endothelial cells (VEC) from bone marrow mesenchymal stem cells (BMSCs), and possibility to providing seed cells for vascularization of tissue engineering. METHODS: BMSCs were isolated from posterior limbs of rat and expanded in vitro. BMSCs of the third generation were cultured with the conditional medium for 14 d. The cells were divided into induced group (containing VEGF, bFGF endothelial cell medium) and control group (containing M199 basic medium). The proliferation activity of cells was tested by MTT. The CD34 and CD31 proteins were examined by immunocytochemistry staining and the gene expression of angiogeninl and angiogenin2 were detected by RT-PCR. RESULTS: There was no significant difference in proliferation activity between induced group and control group (P1d = 0.855, P2d = 0.053, P3d = 0.249, P4d = 0.059, P5d = 0.174). The induced cells were positive for the marker CD34 and CD31 proteins , and RT-PCR showed gene expression of mature endothelial-specific markers angiogeninl and angiogenin2. CONCLUSION: BMSCs can be differentiated into cells with characteristics of vascular endothelial cells, they are ideal seed cells of tissue engineering vascularization.
bone marrow mesenchymal stem cellsvascular endothelial cellsinduceddifferentiationPCR
NETL
NSTL
万方数据
维普
