产后小鼠子宫内膜边缘群干/祖细胞的分离纯化

Isolation and purification of side population stem cells from postpartum mouse endometrium

目的分离和纯化小鼠子宫内膜边缘群(side population,SP)干/祖细胞,为进一步探讨产后子宫修复的细胞机制奠定基础。方法分别采用酶消化和机械分离结合法与机械研磨分离的方法,原代分离小鼠子宫内膜上皮细胞与基质细胞,通过Hoechst33342-SP法(对照加入维拉帕米),使用流式细胞仪检测子宫内膜SP细胞。结果未孕小鼠子宫内膜上皮与基质中均无明确SP干/祖细胞,产后哺乳期后的小鼠子宫内膜基质中含有SP干/祖细胞达(3.44±1.59)%。结论本研究初步检测到小鼠产后子宫内膜中含有SP干/祖细胞。产后更多SP细胞的出现可能与修复损伤的子宫内膜有关。这群SP细胞是子宫原位干细胞还是有其他来源,如何参与子宫内膜的修复,其分子机制如何,尚待进一步研究。

Objectives： To isolate and purify side population（SP） stem/progenitor cells in postpartum mouse endometrium.Methods： Enzymatic digestion combined with mechanical isolation was used to isolate endometrial epithelial and stromal cells from virgin or postpartum mice.The cells were divided into two aliquots.One aliquot was incubated with Hoechst 33342 dye only,and the others with Hoechst in the presence of verapamil as control.Flow cytometry was used to detect the SP cells.Results： No SP stem/progenitor cells were detected in endometrial epithelial or stromal cells of virgin mice and the epithelial cells from postpartum mice did not contain SP stem/progenitor cells either,while SP stem/progenitor cells（3.44%±1.59%） of endometrial stromal cells from postpartum mice were detected. Conclusions： Our investigation found the existence of SP stem/progenitor cells in mouse endometrium.We propose that emergence of more stem cells in the endometrium after delivery could be involved in the repairing of the damaged endometrium.Whether these SP cells originate from uterine itself or other tissues and how they participate in endometrium repair,deserve further study.