人早期胚胎间充质干细胞的定位和体外初步分离培养  

In vitro isolation and cultivation of human fetal mesenchymal stem cells

作　　者：仵敏娟 [1] 刘善荣 [1] 刘厚奇 [2] 惠宁 [1] 杨玲 [1] 王凤玫 [1] 汤淑萍 [1] 

机构地区：[1]第二军医大学基础医学部组织胚胎学教研室,上海,200433 [2]长海医院妇产科,上海,200433

出　　处：《第二军医大学学报》2004年第8期818-821,共4页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(90208026).

摘　　要：目的:从早期人胚分离培养人早期胚胎间充质干细胞(mesenchymal stem cell,MSC),并初步鉴定其生物学特性。方法:取4-6周胚龄的人胚,用免疫组织化学法,结合特定抗体SH-2,对MSC在人胚中的分布进行定位,分离组织进行培养。免疫组化和流式细胞术检测MSC的相对特异性抗原SH-2、CD44、ODT-4、S-100、CD34、α-smooth-actin在分离培养细胞中的表达。结果:在胚胎的四肢、躯体部紧邻神经管的下方(不包括原肠等内脏部位)有大量的MSC:分布。分离培养可得到成纤维状的MSC,贴壁生长,有生长优势,可多次传代并保持MSC:生物学特性不变。结论:通过机械分离和胰酶消化的方法,可从早期人胚得到MSC。利用MSC的贴壁生长特性及生长优势,经4-5次传代后可得到纯度很高的MSC。Objective:To isolate and cultivate human fetal mesenchymal stem cells(MSCs) and to study their biological characteristics. Methods: Human MSCs distribution were detected by histological staining with specific antibody SH-2 at different stages of human embryo from 4 to 6 weeks, then the tissues were isolated and cultured in vitro. The expressions of SH-2, CD44,OCT-4,S-100,CD34, and α-smooth-actin in the cultured cells were detected by immunochemistry and flow cy tometry(FCM). Results: MSCs mainly located in the limb bud and under the neuratube (not include primitive gut or hepatic bud). The fibroblast-like cells were isolated and cultured, which had the ability to proliferate and adhere. The MSCs were passaged many times without significant biological changes. Conclusion: Human fetal MSCs can be obtained from human early fetus by mechanical isolations and trypsinizations. After passaged for 4-5 times the MSCs are very pure.

关 键 词：间充质干细胞 胚胎 细胞分离 细胞培养 

分 类 号：Q2-33[生物学—细胞生物学] Q813.11