中国组织工程研究与临床康复2009，Vol.13Issue(10)：1996-2000,5.
微载体cytodex3培养技术在大量扩增成人骨髓间充质干细胞中的应用
Microcarrier cytodex3 culture technique for amplification of a large amou nt of adult bone marrow mesenchymal stem CellS
摘要
Abstract
OBJECTIVE:To create an in vitro harvesting method of culturing a large number of adult bone marrow MSCs(BMSCs) DESIGN,TIME AND SETTlNG:The randomized, controlled study was performed at the Shanghai Institute of Digestive Surgery (Key Laboratory of Education Committee of Shanghai City),as well as Department of General Surgery and Organ Transplantation Center,Ruijin Hospital,Medical College.Shanghai Jiao Tong University from September 2005 to April 2006.MATERIALS:Bone marrow samples were collected from normal persons.who did bone marrow examination at the Department of Hematology,Ruijin Hospital,Medical College.Shanghai Jiao Tong University.Donors were volunteers who signed the informed consent.METHODS:Human BMSCs were harvested using Pemoll gradient centrifugation and adherence method.and then incubated in microcarrier cytodex3.Common monolayer polystyrene was incubated as controls.Cell phenotype and proliferative activity were tested utilizing flow cytometry and MTT.MAIN OUTCOME MEASURES:Collection.incubation,morphology of human BMSCs.and prolireration and cell cycle of human BMSCs on the cytodex 3 were measured.RESULlTS:Flow cytometry detection showed that the surface marker of human BMSCs on the cytodex3 was ldentical to that on the common monolayer polystyrene;BMSCs were positive for CD29,CD44 and CD105.but negative for CD14,CD34,CD45,VLA-1 and HLA-DR.MTT detection demonstrated that human BMSCs were in the adaptive phase at days 1-3.and entered logarithmic phase frOm day 3.No significant difference was detected in human BMSCs on the monolayer polystyrene and cytodex3(P＞0.05).On the monolayer polystyrene,human BMSCs entered degenerating stage from day 6,whereas on the cytodex3,human BMSCs were still in the logarithmic growth phase at day 9(P＜0.05).Flow cytometry detection confirmed that the cell cycle of human BMSCs was the same both on the monolayer polystyrene and cytodex3 (P＞0.05). CONCLUSION:Using cytodex3 culture technique,a large amount of human BMSCs can be obtained,and the proliferative activity of these BMSCs is good.关键词
间充质干细胞/细胞培养/微载体分类
医药卫生引用本文复制引用
韩宝三,沈柏用,张瑞,王兆海,杜志勇,程东峰,施敏敏,李宏为,彭承宏..微载体cytodex3培养技术在大量扩增成人骨髓间充质干细胞中的应用[J].中国组织工程研究与临床康复,2009,13(10):1996-2000,5.基金项目
国家高新技术研究发展计划-"863"项目:微粒生物人工肝反应器构建的研究,项目编号:2008AA022417 （）
国家自然科学基金:新型微粒生物人工胰反应器构建的研究,项目编号:30801107 （）
国家自然科学基金:微粒肝细胞生物反应器-微囊化三维多孔微载体构建的研究,项目编号:30672043 （）
国家自然科学基金:新型超分子自组装微囊,肝细胞球形聚集体构建的研究,项目编号:30772105 （）
国家自然科学基金:人肝细胞,聚合物杂化界面和生物人工肝的研究,项目编号:20074031 （）
上海市科委重点项目:活体肝移植的应用基础研究,项目编号:07JC14040 （）
上海市科委自然科学基金:超分子自组装可控半透膜/肝细胞球形聚集体构建的研究,项目编号:07ZR14076 （）
上海市教委科研创新项目:新型超分子自组装半透膜微囊胰岛移植的实验研究,项目编号:09YZ84 （）
山东省科技攻关项目:肝细胞微载体构建的研究,项目编号:2008GG100002027. （）
