茶多酚对人脂肪来源间充质干细胞成骨分化的影响  

Effect of EGCG on osteogenic differentiation of human adipose-derived mesenchymal stem cells

作　　者：王华[1] 齐玉成 杨云芳[1] 赵艺洋 王慧 陈培[1] 杨旭芳[1] WANG Hua(Mudanjiang Medical University,Mudanjiang 157011,China)

机构地区：[1]牡丹江医学院,黑龙江牡丹江157011 [2]南方医科大学第一临床医学院,广东广州510515

出　　处：《牡丹江医学院学报》2021年第1期1-3,58,共4页Journal of Mudanjiang Medical University

基　　金：牡丹江医学院研究生创新科研项目(2019YJSCX-20MY)。

摘　　要：目的探讨茶多酚(Epigallocatechin-3-gallate,EGCG)对人脂肪间充质干细胞(human adipose-derived mesenchymal stem cells,hADSCs)成骨分化的影响。方法利用胶原酶消化法和贴壁筛选法从人脂肪组织中分离、培养及扩增hADSCs,倒置显微镜下观察各代hADSCs的形态学特点;利用流式细胞术检测各代hADSCs免疫学表型;取P3代细胞进行成骨诱导分化,实验分三组,即未诱导组、常规成骨诱导组与EGCG组(常规成骨诱导+5μmol/L EGCG),14 d后,镜下观察细胞形态学改变及碱性磷酸酶(ALP)染色。结果体外分离、培养的hADSCs形态均一;流式细胞术结果显示hADSCs具备间充质干细胞的免疫学表型,即CD44、CD73、CD105阳性;成骨诱导14 d后部分细胞由长梭形变成多角形,细胞呈现聚集趋势;ALP染色显示EGCG组呈强阳性。结论成功的从脂肪组织中分离培养出了hADSCs,EGCG能加强其成骨分化能力,这将为骨质疏松症的临床药物开发提供新的思路,亦为组织工程骨的构建提供丰富可靠的种子细胞来源。Objective To explore the effect of tea polyphenol EGCG on the osteogenic differentiation of human adipose-derived mesenchymal stem cells(hADSCs).Methods To isolate,culture and amplify hADSCs from human adipose tissue by collagenase digestion and adherent screening methods,the morphological characteristics of each passage of hADSCs were observed under an inverted microscope.The immunophenotype of each generation of hADSCs was detected by flow cytometry.P3 passage cells were taken for osteogenic induction and differentiation,and were divided into three groups:non-induced group,conventional osteogenic induction group and EGCG group(conventional osteogenic induction with+5μmol/L EGCG).After 14 days,morphological changes and alkaline phosphatase(ALP)staining were observed under the microscope.Results The morphology of hADSCs isolated and cultured in vitro was uniform.The results of flow cytometry showed that hADSCs had the immunophenotype of mesenchymal stem cells,such as CD44,CD73 and CD105.After 14 days of osteogenic induction,some cells changed from long spindle shape to polygonal shape,and the cells showed aggregation trend.ALP staining showed strong positive in EGCG group.Conclusion hADSCs have been successfully isolated and cultured from adipose tissue.EGCG can enhance the osteogenic differentiation ability of hADSCs,which will provide a new idea for the clinical drug development of osteoporosis and provide an abundant and reliable source of seed cells for the construction of tissue-engineered bone.

关 键 词：EGCG 人脂肪来源间充质干细胞 成骨分化 

分 类 号：R595.2[医药卫生—内科学]