不同培养方法对人脐带间充质干细胞的影响  

Different culture methods for human umbilical cord mesenchymal stem cells

作　　者：刘奉[1] 蒋祥林[1] 刘明明[2] 

机构地区：[1]重庆三峡医药高等专科学校,重庆市404020 [2]重庆医科大学基础医学院,重庆市400016

出　　处：《中国组织工程研究》2016年第28期4136-4141,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:脐带干细胞培养主要来源于足月儿,常用培养方法包括组织贴块法、胰酶消化法,但是对于不同培养方法对脐带间充质干细胞的分离结果尚存在较大的争议。目的:观察不同培养方法对脐带间充质干细胞的影响。方法:取30例足月妊娠剖宫产健康胎儿脐带,采用组织贴壁法和胶原酶-胰酶消化法进行培养,制备细胞悬液,获得人脐带间充质干细胞进行分离培养,在流式细胞仪上测定细胞生长情况。结果与结论:(1)细胞融合情况:脐带经过组织贴壁法原代培养5 d后,细胞开始从脐带组织分离,呈现梭形,培养10 d后细胞融合达到80%,脐带经胶原酶-胰酶消化法培养5 d后,可见少量形态各异的贴壁细胞分布,纤维样细胞培养2周后培养融合才达到80%;(2)细胞生长情况:两种不同培养方法分离出的脐带间充质干细胞生长情况差异无显著性意义(P>0.05);(3)细胞鉴定结果:两种培养方法培养的细胞CD13,CD29,CD44及CD105标志物表达均为阳性。(4)结果说明:人脐带间充质干细胞采用组织贴壁原代培养法培养成功率高,优于胶原酶-胰酶消化法培养。BACKGROUND：Umbilical cord stem cel s mainly derive from ful-term infants, and common culture methods include tissue-attached method and trypsin-digestion mehod. However, effects of different culture methods on the separation of umbilical cord mesenchymal stem cel s remain many disputes. OBJECTIVE：To observe the effects of different culture methods on umbilical cord mesenchymal stem cel s. METHODS：Umbilical cords of 30 healthy ful-term and caesarean delivery infants were selected, and cultured using tissue-attached method or trypsin-digestion method to isloate and culture human umbilical cord mesenchymal stem cel s. Meanwhile, cel growth was measured by flow cytometry. RESULTS AND CONCLUSION：The fusiform-shaped cel s began to separate from the umbilical cord tissue that was primary cultured using tissue-attached method, and 10 days later, the cel fusion reached 80%;after the umbilical cord was cultured using col agenase-trypsin digestion for 5 days, a smal amount of adherent cel s with different shapes appeared, and the fiber-like cel s reached 80%of confluence until 2-week culture. There was no significant difference in the growth of umbilical cord mesenchymal stem cel s cultured by different culture methods （P〉0.05）. Moreover, cel s cultured by two methods were al positive for CD13, CD29, CD44 and CD105. These results demonstrate that human umbilical cord mesenchymal stem cel s exhibit a high success rate in primary culture using tissue-attached method, which is superior to the trypsin-digestion method.

关 键 词：组织工程 脐带 干细胞 脐带脐血干细胞 培养方法 脐带间充质干细胞 剖宫产 组织贴块法 胰酶消化法 培养瓶 造血干细胞 

分 类 号：R394.2[医药卫生—医学遗传学]