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胎儿肝脏与骨髓间充质干细胞的分离鉴定

王博蔚1,刘志辉2,刘爱民1,韩丽英1,李荷莲1*   

  1. 1. 吉林大学第二医院妇产科,吉林 长春130041;2. 吉林大学口腔医院修复科,吉林 长春130041
  • 收稿日期:2005-06-06 修回日期:1900-01-01 出版日期:2006-01-28 发布日期:2006-01-28
  • 通讯作者: 李荷莲

Isolation and identification of mesenchymal stemcells from fetal liver and bone marrow

WANG Bo-wei1, LIU Zhi-hui2, LIU Ai-min1, HAN Li-ying1, LI He-lian1*   

  1. 1. Department of Obstetrics and Gynecology, Second Hospital,Jilin University, Changchun 130041, China; 2. Department of Renovation, Stomatology Hospital, Jilin University,Changchun 130041, China
  • Received:2005-06-06 Revised:1900-01-01 Online:2006-01-28 Published:2006-01-28
  • Contact: LI He-lian

摘要: 目的:建立胎儿肝脏与骨髓间充质干细胞(MSCs) 的分离培养方法,并对两种来源细胞的生物学特性加以鉴定。方法:取胎龄4或5月水囊引产胎儿肝脏和骨髓,用1.073 g·mL-1的percoll密度梯度离心分离低密度细胞,MSCs培养基纯化贴壁细胞,用流式细胞术检测其表面标志,成脂肪及成骨诱导鉴定多向分化潜能,对比两种来源的MSCs增殖及生长特征。 结果:胎儿肝脏及胎儿骨髓MSCs CD29、CD44、CD73(SH-3,4)、CD105(SH-2)、CD166、HLA-ABC阳性,CD35、CD45、HLA-DR阴性,均能成脂肪及成骨诱导分化,随MSCs 数量增加,其对外周血T 淋巴细胞转化的抑制作用增强。结论:胎儿肝脏和骨髓中可成功分离MSCs。

关键词: 胎儿, 肝脏, 骨髓, 细胞分离

Abstract: Objective To isolate and identify the phenotype and biological characteristics of fetal liver derived mesenchymal stem cells(MSCs) as well as the MSCs derived from fetal bone marrow. Methods Mononuclear cells from fetal liver were collected by Percoll gradient centrifugation. The low density cells were cultivated and expanded in MSCs media. Phenotype was analyzed by FACS flow cytometer and characteristics of the multipotent were observed. Results The expressions of CD29,CD44,CD73(SH-3,4),CD105(SH -2),CD166,HLA-ABC in MSCs from fetal liver and fetal bone marrow were positive,CD35,CD45,HLA-DR were negative. MSCs from fetal liver and bone marrow were successfully cultivated and differentiated into osteoblast and adipose in vitro. As the quantity of MSCs increased,their inhibitory effects on transformation of lymphocytes in peripheral blood enhanced. Conclusion MSCs existing in fetal liver and bone marrow can be isolated successfully.

Key words: fetal, liver, bone marrow, cell separation

中图分类号: 

  • Q813.11