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Differentiation of side population cells isolated from human amniotic mesenchymal cells into vascular endothelial cells

机译:从人羊膜间充质细胞分离的侧群细胞分化为血管内皮细胞

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Human amniotic mesenchymal side polulation (SP) cells have pluripotency and weak immunogenicity. We attempted to clarify appropriate condition on differentiation of the SP cells into vascular endothelial lineage and to construct tubular formation. The human amniotic mesenchymal cells were isolated from the enzyme-treated amniotic membrane procured from caesarean section and stained with Hoechst33342. SP cells were negatively selected by sorting with fluorescence-activated cell sorting. The expression levels of vascular endothelial markers were evaluated by real-time PCR and immunostaining. The expression levels of HLA class I and class II genes were evaluated by using microarray. Vascular endothelial markers like vWF and KDR in SP cells were more strongly induced by vascular endothelial growth factor (VEGF) in hypoxia for 2 weeks, but long-term induction under hypoxia was not effective for amniotic mesenchymal SP cells to differentiate into vascular endothelial cells. Besides, human amniotic mesenchymal SP cells constructed tubular formation on the Matrigel~(TM), indicating that the cells had angiogenic potential.
机译:人羊膜间充质侧细胞(SP)细胞具有多能性和弱免疫原性。我们试图阐明将SP细胞分化为血管内皮谱系的适当条件,并构建小管形成。从剖腹产后经酶处理的羊膜中分离人羊膜间充质细胞,并用Hoechst33342染色。通过荧光激活细胞分选法分选阴性的SP细胞。通过实时PCR和免疫染色评估血管内皮标志物的表达水平。通过使用微阵列评估HLA I类和II类基因的表达水平。缺氧持续2周,血管内皮生长因子(VEGF)可以更强烈地诱导SP细胞中的血管内皮标记物vWF和KDR,但缺氧条件下的长期诱导对羊膜间充质SP细胞分化为血管内皮细胞无效。此外,人羊膜间充质SP细胞在Matrigel TM上形成管状形成,表明该细胞具有血管生成潜能。

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