中国组织工程研究Issue(23)：3609-3615,7.DOI:10.3969/j.issn.2095-4344.2014.23.001
Notch信号通路阻断剂对人脐带间充质干细胞软骨分化的影响
A Notch signaling pathway inhibitor affects chondrogenesis of human umbilical cord mesenchymal stem cells
摘要
Abstract
BACKGROUND:Notch singling pathway is very important for cellproliferation and differentiation, but its role is stil unknown during chondrogenesis of human umbilical cord mesenchymal stem cells. OBJECTIVE:To investigate the effect of N-[N-(3,5-difluorophenacetyl-L-alanyl)]-(S)-phenylglycinet-butyl ester (DAPT) on inducing human umbilical cord mesenchymal stem celldifferentiation into chondrocytes. METHODS:Human umbilical cord mesenchymal stem cells were isolated from human umbilical cord, then were induced to differentiate into chondrocytes. There were four experimental groups:non-induced group, high-glucose Dulbecco’s modified Eagle’s medium containing 5%fetal bovine serum and 1%double antibody;induced group, induced medium containing 6.25 mg/L insulin, 6.25 mg/L transferrin, 10μg/L transforming growth factor beta 1, 0.1μmol/L dexamethasone, 50 mg/L vitamin C, 5%fetal bovine serum and 1%double antibody;dimethyl sulfoxide group, induced medium containing 0.1%dimethyl sulfoxide;DAPT group, induced medium containing 5μmol/L DAPT. RESULTS AND CONCLUSION:After chondrogenic induction, the morphology of human umbilical cord mesenchymal stem cells became polygon and positive for toluidine blue and immunofluorescence staining;the expression of Jag-1, PS-1, Notch-1 and Hes-1 decreased significantly (P<0.01). After the addition of DAPT, compared with the induced group, the relative gene expression of Jag-1, PS-1 and Hes-1 decreased markedly (P<0.01), the relative gene expression of Notch-1 decreased obviously as wel (P<0.05), and the contents of proteoglycan and col agen type II proteins decreased significantly (P<0.01). At the same time, the relative gene expression of proteoglycan decreased obviously (P<0.05), and the relative gene expression of col agen type II decreased in part. Notch signaling pathway exists in human umbilical cord mesenchymal stem cells, once chondrogenesis begins, the signaling strength wil decline rapidly. DAPT may prevent human umbilical cord mesenchymal stem cells from differentiating into chondrocytes by Jag-1-Notch-1-Hes-1 pathway.关键词
干细胞/脐带脐血干细胞/分化/人脐带间充质干细胞/软骨细胞/DAPT/Notch信号通路/蛋白聚糖/Ⅱ型胶原蛋白/甲苯胺蓝染色/免疫荧光染色/qPCR/广东省自然科学基金Key words
stem cells/mesenchymal stem cells/umbilical cord/chondrocytes/receptors, Notch分类
医药卫生引用本文复制引用
张家文,罗二梅,吴颜晖,符淑莹,陈丹纯,宇丽,唐明乔..Notch信号通路阻断剂对人脐带间充质干细胞软骨分化的影响[J].中国组织工程研究,2014,(23):3609-3615,7.基金项目
广东省自然科学基金资助项目(9151008901000050)；暨南大学大学生创新创业训练计划(Cx12200)Funding:the Natural Science Foundation of Guangdong Province, No.9151008901000050 （9151008901000050）
Innovation and Entrepreneurship Training Program for Students in Jinan University, No. Cx12200 （）
