子宫内膜基质干细胞的体外成骨诱导和成脂诱导分化

目的探讨子宫内膜基质干细胞的多向分化潜能。方法取因子宫肌瘤行全子宫切除术患者的子宫内膜组织,磁珠分选子宫内膜基质干细胞,进行传代培养。传代细胞分别加入成骨诱导剂和成脂诱导剂培养,并通过茜素红和油红O染色对成骨细胞和脂肪细胞形态进行鉴定。结果子宫内膜基质干细胞呈成纤维细胞样贴壁生长,其经成骨、成脂诱导培养3周后形态、体积发生明显改变。茜素红染色显示细胞团中央能形成钙化结节;成脂诱导后油红O染色可见细胞质内出现橙红色脂滴。结论子宫内膜基质干细胞经体外诱导培养后可向成骨细胞和脂肪细胞分化,并具有明显的成骨和成脂细胞形态,表明子宫内膜基质干细胞具有多向分化潜能。

Differentiation of endometrial stromal stem cells into osteoblasts and adipocytes in vitro

Objective To investigate the multilineage differentiation capacity of stromal stem cells isolated and cultured from human endometrial tissues.Methods Single-cell suspensions of endometrial stromal stem cells were obtained from hysterectomy tissues of women experiencing normal menstrual cycles.Purified stromal stem cell suspensions were then obtained by selecting cells with a further round of magnetic bead sorting using anti-EpCAM-coated Dynabeads.Rare human endometrial EpCAM-stromal stem cells were screened by inverted microscope each day and identified by flow cytometer.Then the cells were cultured in osteoblast-inducing culture medium,and osteoblast phenotype was assayed with alizarin red staining.The passage cells were cultured in adipogenesis-medium and stained with oil red O for identification.Results Endometrial stromal stem cells grew as adherent cells and presented fibroblast-like in vitro,and could stably proliferate and be passed.The cells changed from fibroblast-like into ellipse after osteoblast-inducing cultivation.After induction for 21 d,alizarin red staining demonstrated the formation of mineralized nods in extracellular matrix.Under the inverted microscope,there existed significant lipid drops around the cell nucleus after adipogenesis-inducing cultivation for 21 d and detected by oil red O.Conclusion Endometrial stromal stem cells can be induced to be osteo-inducing and adipogenisic,suggesting that the cells have the multilineage differentiation.