低氧条件对小鼠ADSCs膜片形成及其生物学特性的影响

The effects of hypoxia on the formation and biological properties of mouse adipose-derived stem cell sheet

目的:观察低氧条件对小鼠脂肪间充质干细胞(ADSCs)膜片生物学性能的影响。方法:分离培养C57/BL小鼠ADSCs并用Vit C法构建细胞膜片;氯化钴(CoCl_2)模拟体内低氧环境,q PCR和Western Blot检测HIF-1α表达;组织学染色、透射电镜观察膜片形成;q PCR检测ADSCs干性及膜片分泌功能相关基因的表达。结果:成功构建ADSCs膜片;HIF-1α的表达随CoCl_2浓度的升高而升高;低氧条件下,形成细胞膜片较薄,但膜片内细胞连接更紧密;低氧环境可增强ADSCs干性,促使ADSCs膜片合成更多黏附分子、分泌更多保护性生长因子。结论:低氧条件更利于ADSCs膜片构建、存活和生物学功能维持。

Objective：To study the effects of hypoxia on the formation and function of mouse adipose-derived stem cell（ADSC） sheet.Methods：The ADSCs were isolated from 8-weeks-old C57/BL mouse visceral adipose tissue and cultured with Vit C to get cell sheets.The sheets were observed under inverted microscope.CoCl2 was added in the culture system to simulate the hypoxia environment,which was verified by the expression of hypoxia inducible factor-1α（HIF-1α） through qPCR.Transmission electron microscope observation,histological staining and qPCR were conducted to analyze the influences of hypoxia on ADSCs in terms of sheet-formation,stemness property and secretion function.Results：The ADSC sheets were obtained successfully.CoCl2 showed hypoxia-inductive ability with a corresponding increase of HIF-1α expression.The hypoxia condition produced thinner cell sheets with less collagenⅠin the extracellular matrix,lead to more cell-to-cell connection and maintainess of the stemness property of ADSCs and promoted the secretion of adhesion molecules and protective growth factors with the cell-sheet formation.Conclusion：Hypoxia environment can promote ADSCs cell sheet formation,survival and biological function.