广西医科大学学报2019，Vol.36Issue(4)：538-542,5.DOI:10.16190/j.cnki.45-1211/r.2019.04.010
试剂盒法与差速超速离心法所提取的人牙髓细胞外泌体的鉴定与比较
Identification and comparison of human dental pulp cells derived exosomes extracted by ExoQuick kit and ultra-centrifugation
摘要
Abstract
Objective:To identify and compare human dental pulp cells derived exosomes(hDPC-DEs) extracted by ExoQuick kit and ultra-centrifugation.Methods:Human dental pulp cells(hDPCs) were primarily cultured and identified,and the hDPC-DEs were extracted by ExoQuick kit or ultra-centrifugation.The morphology of extracted exosomes was observed under transmission electron microscopy.The expression of exosomal marker protein CD9was detected by western blotting,and the concentration of exosomal marker protein CD63was determined by ELISA.Results:HDPC-DEs obtained by ExoQuick kit and ultra-centrifugation methods showed typical saucer-like double-layer membrane structure,with the diameter of about 50-120nm under electronic microscopy.There were more impurities in ExoQuick kit group.The exosomal marker protein CD9in ExoQuick kit and ultra-centrifugation groups were positively expressed,and the concentration of the exosomal marker protein CD63 in the ExoQuick kit group was significantly higher than that in the ultra-centrifugation group(P<0.05).Conclusion:The hDPC-DEs could be successfully extracted by both ultra-centrifugation and ExoQuick kit.The latter method was easy to operate,and could obtain higher concentration of hDPC-DEs,but more impurities.关键词
人牙髓细胞/外泌体/试剂盒法/差速超速离心法Key words
human dental pulp cells/exosome/ExoQuick kit/ultra-centrifugation分类
医药卫生引用本文复制引用
郑程峰,陈硕琳,陈文瑨,林万惠,吴周玲,韦慧妮,陈文霞..试剂盒法与差速超速离心法所提取的人牙髓细胞外泌体的鉴定与比较[J].广西医科大学学报,2019,36(4):538-542,5.基金项目
国家自然科学基金资助项目(No.81760195) （No.81760195）
