中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (23): 4233-4236.doi: 10.3969/j.issn.1673-8225.2011.23.013

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

人胚胎干细胞饲养层的制备及生物学活性

胡嘉波，麻全慧，胡三强，周中卫，王晓慧，钱  晖，许文荣   

  1. 江苏大学基础医学与医学技术学院，江苏省镇江市  212001
  • 收稿日期:2010-11-30 修回日期:2011-03-16 出版日期:2011-06-04 发布日期:2011-06-04
  • 通讯作者: 许文荣，博士，教授，江苏大学基础医学与医学技术学院，江苏省镇江市 212013 icls@ujs.edu.cn
  • 作者简介:胡嘉波☆，男，1969年生，江苏省如东县人，汉族，2008年江苏大学毕业，博士，副教授，主要从事干细胞方面的研究。 hu@ujs.edu.cn
  • 基金资助:

    江苏大学高级人才专项资助(09JDG037)；国家自然科学基金资助项目(31071421)。

Preparation and biological characteristics of feeder cells for human embryonic stem cells

Hu Jia-bo, Ma Quan-hui, Hu San-qiang, Zhou Zhong-wei, Wang Xiao-hui, Qian Hui, Xu Wen-rong   

  1. School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang  212001, Jiangsu Province, China
  • Received:2010-11-30 Revised:2011-03-16 Online:2011-06-04 Published:2011-06-04
  • Contact: Xu Wen-rong, Doctor, Professor, School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang 212001, Jiangsu Province, China icls@ujs.edu.cn
  • About author:Hu Jia-bo☆, Doctor, Associate professor, School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang 212001, Jiangsu Province, China hu@ujs.edu.cn
  • Supported by:

    Startup Foundation for Advanced Talents, Jiangsu University, No. 09JDG037*; the National Natural Science Foundation of China, No.31071421*

PDF

354

被引次数

14

摘要：

背景：建立一种既可以大量制备，又能保存并保持较高活性的饲养层是胚胎干细胞培养研究不可缺少的环节。
目的：体外分离培养、冻存复苏ICR小鼠胚胎成纤维细胞，观察其生物学特性。
方法：取ICR小鼠13.5 d胚胎，用胰蛋白酶分步消化法分离培养小鼠成纤维细胞，对冻存复苏后的小鼠胚胎成纤维细胞形态、生长曲线、贴壁率、细胞化学染色及支持人胚胎干细胞生长特性等进行观察。
结果与结论：复苏后的小鼠胚胎成纤维细胞在体外传代30 min时80%以上细胞贴壁，生长曲线显示细胞增殖活跃，细胞化学染色AKP、PAS、POX阴性，能长期支持人胚胎干细胞传代生长。提示此方法所获得的小鼠胚胎成纤维细胞复苏后有较高的生物学活性，可为人胚胎干细胞扩增提供稳定、优质的饲养层细胞。

关键词: 小鼠胚胎成纤维细胞, ICR小鼠, 人胚胎干细胞, 冻存, 复苏

Abstract:

BACKGROUND: It is important to establish and save a large number of high-activity feeder cells for the culture of human embryonic stem cells (hESCs).
OBJECTIVE: To separate, culture, freeze and thaw embryonic fibroblasts of ICR mice in vitro, and to explore the biological characteristics of mouse embryonic fibroblast.
METHODS: ICR mouse fetuses of 13.5 days were chosen to isolate mouse embryonic fibroblasts by trypsin stepwise. We observed the cell morphology, growth curves, cells adhesive rate, and cytochemistry staining of mouse embryonic fibroblasts after cryopreservation. The effect of mouse embryonic fibroblasts was assessed by the ability to support the growth of hESCs.
RESULTS AND CONCLUSION: Thirty minutes after passage, the adhesive rate of mouse embryonic fibroblasts was above 80%. Mouse embryonic fibroblasts proliferated quickly. AKP, POX and PAS cytochemistry staining of mouse embryonic fibroblasts were all negative, indicating that it could support hESCs in a long period. The thawed mouse embryonic fibroblasts have high biological activity, and can be used as the stable and high quality feeder cells for hESCs culture.

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