脱水淫羊藿素维持人尿源性干细胞干性的作用研究

目的:探究脱水淫羊藿素(DICT)对人尿源性干细胞(hUSCs)干性的影响。方法:常温离心法体外分离培养hUSCs,倒置相差显微镜下进行hUSCs形态学观察;根据MTT实验绘制hUSCs接种后1~7 d的生长曲线;流式细胞术检测hUSCs表面标志物;分别设hUSCs培养基对照组,浓度为0.5、1.0、1.5、2.0、2.5、3.0μmol/L的DICT处理组,利用四甲基噻唑蓝(MTT)实验检测作用1~7 d后各组的细胞活力;采用实时荧光定量PCR(qPCR)检测2.0μmol/L DICT作用hUSCs 3 d后,对干性基因OCT-4、C-MYC mRNA表达的影响。结果:通过常温离心法成功获得形态良好、生长活性较高的hUSCs,hUSCs表达间充质干细胞表面标志物CD44和CD90,不表达造血干细胞表面标志物CD34和内皮细胞表面标志物CD31。2.0μmol/L DICT作用3~6 d能显著促进hUSCs的增殖,增强其生长活力(P<0.01),上调hUSCs干性基因OCT-4、C-MYC的mRNA表达(P<0.01)。结论:DICT在体外可增强hUSCs的干性。

Usefulness of dehydrated icaritin in maintaining the stemness of human urine-derived stem cells

OBJECTIVE:To investigate effects of dehydrated icaritin (DICT) on the stemness of human urine-derived stem cells (hUSCs). METHODS:hUSCs were isolated by centrifugation at room temperature and cultured in vitro. Morphological observations of hUSCs was performed under inverted phase contrast microscope. The growth curve of hUSCs from 1-7 days was plotted according to MTT experiment. The surface markers of hUSCs were detected by flow cytometry. The control group of hUSCs culture medium and DICT treatment groups with concentrations of 0.5,1.0,1.5,2.0,2.5,3.0 μmol/L were set up. Cell viability of each group was measured by the tetramethylthiazole blue (MTT) assay from 1-7 days. Effects of 2.0 μmol/L DICT treatment of hUSCs on the expression of OCT-4 and C-MYC mRNA were measured by quantitative real-time PCR (qPCR) after 3 days. RESULTS:hUSCs with good morphology and high growth activity were successfully obtained. hUSCs expressed CD44 and CD90,surface markers of MSCs,but not CD34,surface marker of hematopoietic stem cells,and CD31,surface marker of endothelial cells. 2.0 μmol/L DICT treatment for 3-6 days significantly promoted proliferations of hUSCs,enhanced their growth viability (P<0.01),and up-regulated the mRNA expression of hUSCs stemness genes OCT-4 and C-MYC (P<0.01). CONCLUSION:DICT enhanced the stemness of hUSCs in vitro.