人脐带间充质干细胞的分离鉴定及其修复缺血缺氧脑损伤作用  

Isolation and identification of human umbilical cord mesenchymal stem cells and their repair effects on hypoxic-ischemic brain damage

作　　者：江伟[1,2] 沈洁[3] 史源[3] 龚敏[1,2] 瞿平[1,2] 张赟[1,2] 魏小平[1,2] 刘友学[1,2] 陈洁[1,2] 李廷玉[1,2] 

机构地区：[1]重庆医科大学附属儿童医院儿童营养中心,重庆400014 [2]重庆医科大学儿童发育疾病省部共建教育部重点实验室,重庆400014 [3]第三军医大学大坪医院儿科,重庆400037

出　　处：《吉林大学学报（医学版）》2011年第2期215-219,I0002,共6页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金重点项目资助课题(30830106)

摘　　要：目的:研究人脐带间充质干细胞(UCMSCs)多向分化潜能,了解UCMSCs移植治疗脑缺血缺氧损伤(HIBD)大鼠的神经修复功能。方法:采用组织消化原代贴壁法分离培养人UCMSCs;取第4和10代UCMSCs进行流式细胞术检测间充质干细胞(MSCs)表面特异标志物表达;染色体分型检测染色体是否畸变;进行成神经、骨、脂诱导,了解其多向分化潜能;制备HIBD大鼠模型,穿梭箱测试对照组、HIBD细胞移植组和HIBD组大鼠的主动逃避率(AARR)和未逃避率(NARR)。结果:流式细胞术结果显示,类似MSCs样细胞强表达CD29、CD44和CD105,极低表达CD34和CD45,符合UCMSCs的特征。多向分化诱导后的细胞染色和免疫荧光结果显示,UCMSCs可诱导分化为神经样细胞、脂肪细胞和成骨细胞,多次传代后仍可多向诱导,且未发生染色体畸变。穿梭箱测试提示,UCMSCs移植组大鼠在测试的第3、4和5天的AARR高于HIBD组(P<0.05)。结论:UCMSCs易获取及纯化,具有多向分化潜能,多次传代功能稳定,可改善脑损伤动物的神经功能。Objective To study the potential for multiple differentiation of human umbilical cord mesenchymal stem cells（UCMSCs） and discuss the repair effects of UCMSCs in treatment of hypoxic-ischemic brain damage（HIBD） in rats.Methods The UCMSCs were isolated and cultured by using tissue digestion and adherent culture;the expressions of mesenchymal stem cells（MSCs） specific markers of the P4 and P10 UCMSCs were detected by flow cytometry,and the aberration condition was judged by chromosomal tests;the potential of multiple differentiation was detected by neurogenic,adipogenic and osteogenic lineages.The active avoidance response rate（AARR） and none avoidance response rate（NARR） were detected by shuttle box test in normal control,abdominal transplantation UCMSCs and HIBD groups.Results The flow cytometry results showed strong expressions of CD29,CD44 and CD105,and very low expressions of CD34 and CD45 in these.The immunofluorescence and cell staining results showed that the UCMSCs could be induced into neural cells,adipocytes and osteoblasts;even after high passages.The UCMSCs could still be differentiated into multiple cells and chromosome aberration did not occur.The shuttle box test results suggested that the AARR in UCMSCs transplantation group was high than that in HIBD group on the 3th,4th and 5th days（P〈0.05）.Conclusion The UCMSCs are easy to be confirmed and purified,and they have multiple differentiation potential,which are still stable with high passages.

关 键 词：人脐带间充质干细胞 多向分化 脑缺血缺氧损伤 染色体分型 穿梭箱 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R651.15[医药卫生—基础医学]