血管组织细胞抽提物体外调控间质干细胞向内皮细胞分化的研究

目的探讨血管组织细胞抽提物对大鼠骨髓间质干细胞(Bone marrow mesenchymal stem cell,MSC)体外培养的影响及向血管内皮细胞分化的能力。方法利用淋巴细胞分离液分离大鼠MSCs,经贴壁法培养扩增后,流式细胞仪检测MSCs表面抗原表达,然后分别以含有大鼠血管组织细胞抽提物、热处理血管组织细胞抽提物以及用磷酸缓冲液代替细胞抽提物的培养基培养,然后分析比较细胞表型变化及检测内皮细胞特异性标志物的表达变化。结果MSCs在含有大鼠血管组织细胞抽提物的培养基中细胞形态没发生显著变化,但是细胞内基因表达变化发生改变,内皮细胞特异性基因CD31、CD34、CD144和eNOS(endothelial nitric oxide synthase,eNOS)高表达。结论血管组织细胞抽提物中含有可诱导大鼠MSCs向内皮细胞分化的物质,说明成体细胞内组份对邻近干细胞的分化命运有着重要的调控作用。

Objective To investigate the differentiation of Mesenchymal stem cell （MSCs） into the endothelial cells by the extracts of the vascular endothelial cells. Methods MSCs were separated from the bone marrow by density gradient centrifugation, wall sticking screening and amplified in vitro. The surface antigens of MSCs were evaluated with the flow cytometry. The 4th passaged cells were induced to differentiate into endothelial cells by means of differentiation induction media containing the extracts of the vascular endothelial cells, the extracts of the vascular endothelial cells treated by the boiling water and the basic medium, respectively. The expression of cell specific genes, including CD31, CD34, CD144 and eNOS, was detected by RT-PCR, and the morphological changes were observed by inverted microscopy. Results It was revealed by flow cytometry that the isolated and cultivated MSCs expressed CD73 and CD90, and did not express CD34. After induction, the morphology of the differentiated has no distinctive changes. But the expression of the endothelial cell specific genes, fourteen days after induction, only was detected in the induced cells with the extracts of the vascular endothelial cells. Conclusion There are some factors in the extracts of the vascular endothelial cells, which can induce the MSCs to differentiate into the endothelial cells. And the fates of stem cells may be regulated by the cellular contents around them.