摘要
目的用体外无血清培养体系分离培养人脐带间充质干细胞。方法取1例足月妊娠剖宫产健康胎儿脐带,采用组织块贴壁培养,使用无血清培养体系培养获得人脐带间充质干细胞。通过形态学观察,成脂、成骨、成软骨诱导分化能力检测及流式细胞术免疫表型检测进行鉴定。结果组织块贴壁培养6天后可见细胞从组织块边沿爬出;培养15天后,达到80%汇合,生长状况良好,细胞呈梭形,为典型的成纤维细胞形态,极性整齐排列,集落呈涡旋状,形态较均一。传代培养的第三代细胞CD29、CD44、CD90、CD105均为阳性表达,不表达CD34和CD45,均具有成脂、成骨、成软骨分化的能力。结论无血清培养体系组织块贴壁法分离培养人脐带间充质干细胞法成功率高,其生物学特性稳定,并能消除由未确定的病原体引起的风险。
Objective To isolate and culture human umbilical cord mesenchymal stem cells in serum-free culture system.Methods Human umbilical cord mesenchymal stem cells(HUC-MSCs)were obtained from the umbilical cord of a healthy fetus who underwent cesarean section in full-term pregnancy by adherent tissue culture and serum-free culture system.Through morphological observation,ability test of adipogenic,osteogenic,chondrogenic induction and immune phenotype detection with flow cytometry to identify cell.Results The fibroblast-like cells began to emerged from the edge of the tissue 6 days later,and after 15 days of culture,80%of the cells converged and grew well.The cells were spindle-shaped.They were typical fibroblasts with neat polarity,and the colonies were spiral and the morphology was relatively uniform.CD29,CD44,CD90 and CD105 were all positive in the third generation cells,but not CD34 and CD45.The cells could be induced to differentiate into adipocytes,osteogenesis,and chondrocytes.Conclusion The method of isolation and culture of human umbilical cord mesenchymal stem cells with no serum culture system and adherent tissue culture has a high success rate,stable biological characteristics,and can eliminate risks caused by unidentified pathogens.
作者
张丽
王文芳
郭芸
邓丹琪
黄雅亮
纪秋琴
张佩莲
ZHANG Li;WANG Wen-fang;GUO Yun;DENG Dan-qi;HUANG Ya-liang;JI Qou-qin;ZHANG Pei-lian(Basical-medical-school of Kunming Medical University,Kunming 650500,China;Department of Dermatology,The Second Affiliated Hospital of Kunming Medical University.Kunming 650101,China)
出处
《皮肤病与性病》
2019年第1期4-6,共3页
Dermatology and Venereology
基金
云南省应用基础研究昆明医科大学联合专项资助(项目号2014FZ028)
关键词
脐带间充质干细胞
分离
贴壁培养
Umbilical cord mesenchymal stem cell
Isolation
Adherent culture
