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目的探讨由胚胎干细胞(embryonic stem cells,ESCs)诱导的内皮细胞(endothelial cells,Ec)与自体胚胎干细胞联合培养时对ESCs成骨作用的影响。方法提取小鼠的胚胎干细胞和胚胎成纤维细胞,诱导胚胎干细胞生成Ec,培养细胞分为四组。A组:胚胎干细胞诱导组;B组:胚胎干细胞诱导组;C组:胚胎干细胞诱导生成的内皮细胞(Ec)诱导组;D组:ESCs和诱导的EC联合培养诱导组。通过干细胞形态的观察、免疫荧光、细胞的增值、碱性磷酸酶以及骨钙素含量的测定从酶学、组织学及生化学等不同方面观测骨髓基质细胞对胚胎干细胞向成骨细胞诱导转化的影响。结果通过细胞免疫荧光染色证实C组培养诱导的细胞为EC。D组ESCs和诱导的Ec联合培养组MTT检测结果各组细胞生长差异没有显著意义(p>0.05),骨钙素和碱性磷酸酶测定D组有明显差异高于其他3组(p<0.05)。结论以胚胎干细胞诱导生成的内皮细胞对胚胎干细胞的成骨有明显的促进作用。
Objective To investigate the effect of embryonic stem cells (ESCs) induced by embryonic stem cells (ESCs) on the osteogenic effect of ESCs. Methods Mouse embryonic stem cells and embryonic fibroblasts were extracted and embryonic stem cells were induced to produce Ec. The cultured cells were divided into four groups. Group A: Embryonic stem cell induction group; Group B: Embryonic stem cell induction group; Group C: Embryonic stem cell induced endothelial cell (Ec) induction group; Group D: ESCs and induced EC co-culture induction group. Through observation of stem cell morphology, immunofluorescence, cell proliferation, determination of alkaline phosphatase and osteocalcin content, the effect of bone marrow stromal cells on the induction of osteoblast transformation from embryonic stem cells was observed from different aspects of enzymology, histology and biochemistry influences. Results C-cell culture-induced cells were confirmed as EC by immunofluorescence staining. There was no significant difference in the MTT results between the group of ESCs and the group of induced Ec (P> 0.05), and the difference between the group of osteocalcin and the alkaline phosphatase of group D was significantly higher than that of the other three groups (p <0.05 ). Conclusion The endothelial cells induced by embryonic stem cells can promote the formation of embryonic stem cells.