人表皮细胞中侧群细胞表型的初步分析

Preliminary analysis of side population phenotypes in human epithelial cells

目的:检测人表皮细胞中是否存在侧群(side population,SP)细胞,并研究该表型细胞是否表达通用干细胞标志物ABCG2和表皮干细胞的标志物整合素α6和β1。方法:运用中性蛋白酶和胰蛋白酶两步消化法从手术切除的人包皮组织中分离获得表皮细胞。经Hoechst33342荧光染料和PI染色,流式细胞仪分析并分选SP细胞。采用流式细胞仪分析SP细胞ABCG2、整合素α6和β1的表达情况。结果:人表皮细胞中存在SP细胞,其比例为0.2%～0.3%。用流式细胞仪可检测到在SP细胞以及总表皮细胞中均有少量细胞表达通用干细胞标志物ABCG2以及表皮干细胞的标志物整合素α6和β1,但二者阳性细胞百分比差异无统计学意义(P>0.05)。结论:表皮细胞中的SP细胞是否是富集的表皮干细胞仍存在疑问,还需要进一步的动物体内移植实验、克隆形成能力和增殖能力的研究证实。

Objective： To explore side population（SP）Cells ih human epithelial cells and to observe the expression of the universal stem cell marker ABCG2 epidermal stem cell markers a, integrin and β, integrin on SP cells. Methods ： Epithelial cells were obtained by digesting human skins with Dispase Ⅱ and Trypsin and stained with Hoechst33342 and PI. The SP cells were analyzed and sorted by the fluorescence-activated cell sorter. Then the expression of ABCG2, α6 integrin and β1 integrin was analyzed by flow cytometry. Results： SP cells accounted for 0.2%-0.3% of total human epithelial cells. Only a small part of cells expressed ABCG2, integrin α6 and integrin β1 of both SP cells and total human epithelial cells detected by flow cytometry .The difference of positive rates between SP cels and total human epithelial cells was not significant. Conclusion： SP cells accounted for 0.2%-0.3% of total human epithelial cells. The difference of positive rates of stem cells marker ABCG2, integrin α 6 and integrin β1 between SP cells and total human epithelial cells was not significant.