不同胎牛血清用于人脐带间充质干细胞培养的比较分析  

Comparison of different fetal bovine serum in cultivation of human umbilical cord stroma-derived mesenchymal stem cells

作　　者：孙伟伟[1] 徐志浩[2] 刘瑞[1] 原志庆[2] 丰慧根[2] 

机构地区：[1]新乡医学院三全学院生命科学技术学院,河南新乡453003 [2]新乡医学院生命科学技术学院,河南新乡453003

出　　处：《新乡医学院学报》2013年第10期787-789,793,共4页Journal of Xinxiang Medical University

基　　金：河南省科技厅自然科学基金项目(编号:122101310100)

摘　　要：目的比较3种不同厂家的胎牛血清用于人脐带间充质干细胞(hUCS-MSCs)培养的效果,探索可靠稳定的hUCS-MSCs培养方法。方法取3种胎牛血清,分别配置成含体积分数10%胎牛血清的培养液。实验分为A、B、C3组,分别使用3种培养液对细胞进行原代培养。将A组细胞分为D、E、F组,分别使用含有3种不同胎牛血清的培养液进行传代培养。流式细胞仪对第3代细胞进行鉴定。结果 A组第7天观察到有细胞迁出,第15天时有细胞迁出的组织块占总数的48%;B组第12天有细胞迁出,第15天时有细胞迁出的组织块占总数的12%;C组第15天有细胞迁出,有细胞迁出的组织块占总数的2%。传代培养至第3代,D、E、F组细胞形态和生长速率未出现明显差异。第3代细胞不表达CD14、CD34、CD45、CD79a和人类白细胞抗原DR分子,表达CD73、CD90。结论 3种胎牛血清用于原代培养hUCS-MSCs的效果有较大差别,A组效果最佳。3种胎牛血清对hUCS-MSCs的传代培养影响不大。Objective To compare the effectiveness of cultivation of human umbilical cord stroma-derived mesenchymal stem cells（hUCS-MSCs） using different brands of fetal bovine serum,and explore a reliable method of cultivation of hUCS-MSCs. Methods Three kinds of fetal bovine serum were dispensed culture solution which containd volume fraction 10%fetal bovine serum.The hUCS-MSCs were divided into group A,B and C and then performed primary culture with three different culture solution respectively.Cells from group A were divided into group D,E and F,and was subcultured by using three different culture solution respectively which contained three different kinds of fetal bovine serum.The third generation cells were assessed by flow cytometer.Results Cells emigration were found on the 7^（th） day in group A and about 48%tissue masses were found to exist cells emigration on the 15^（th） day.Cells emigration were found on the 12^（th） day in group B and about 12%tissue masses were found to exist cells emigration on the 15^（th） day.Cells emigration were found on the 15^（th） day in group C and about 2%tissue masses were found to existing cells emigration on the 15^（th） day.Cells were subcultured to the 3^（rd） generation and there were no obvious difference of cell shape and growth rate among group D,E and F.The third generation cells did not express CD14,CD34,CD45,CD79a and human leukocyte antigen DR（HLA-DR）,but strongly expressed CD73 and CD90.Conclusion The difference of effects of primary culture of hUCS-MSCs using different fetal bovine serums were obvious,and group A get the best effect.Three kinds of fetal bovine serum have little effect on subculture of hUCS-MSCs.

关 键 词：人脐带间充质干细胞 胎牛血清 细胞培养 

分 类 号：R394.2[医药卫生—医学遗传学]