中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (10): 1780-1785.doi: 10.3969/j.issn.1673-8225.2012.10.016

• 胚胎干细胞 embryonic stem cells • 上一篇    下一篇

羊水间充质干细胞体外培养方案的优化及生物学特性***★

王培培1，郑玉宝1，彭  亮1，于  君2，高志良1   

  1. 1中山大学附属第三医院感染科，广东省广州市   510630；2广西医科大学附属医院感染科，广西壮族自治区南宁市 530021
  • 收稿日期:2011-08-01 修回日期:2011-09-01 出版日期:2012-03-04 发布日期:2012-03-04
  • 通讯作者: 高志良，博士，主任医师，博士生导师，主任，中山大学附属第三医院感染科，广东省广州市 510630 gaozl@21cn.com
  • 作者简介:王培培★，女，1986年生，安徽省合肥市人，汉族，中山大学在读硕士，主要从事干细胞对重型肝炎治疗作用的基础研究。 smilewangpeipei@163.com
  • 基金资助:

    国家自然科学基金项目(30971356)；广东省自然科学基金博士启动项目(10451008901004818)；国家“十一五”计划(2008ZX10002-005)：乙型重型病毒性肝炎(肝衰竭)治疗新方案、新方法研究。

Biological characteristics and optimal strategy of human amniotic fluid-derived mesenchymal stem cells cultured in vitro   

Wang Pei-pei1, Zheng Yu-bao1, Peng Liang1, Yu Jun2, Gao Zhi-liang1   

  1. 1Department of Infectious Diseases, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou  510630, Guangdong Province, China; 2Department of Infectious Diseases, Affiliated Hospital of Guangxi Medical University, Nanning  530021, Guangxi Zhuang Autonomous Region, China
  • Received:2011-08-01 Revised:2011-09-01 Online:2012-03-04 Published:2012-03-04
  • Contact: author: Gao Zhi-liang, M.D., Chief physician, Doctoral supervisor, Department of Infectious Diseases, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China gaozl@21cn.com
  • About author:Wang Pei-pei★, Studying for master’s degree, Department of Infectious Diseases, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China smilewangpeipei@163.com
  • Supported by:

    2011-09-01 the National Natural Science Foundation of China, No. 30971356*; the Natural Science Foundation of Guangdong Province, No. 104510089010 04818*; National Eleventh Five-Year Program of China, No. 2008ZX10002-005*

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摘要：

背景：目前干细胞工程种子来源多样化，羊水中存在胎儿脱落细胞，可分离培养出间充质来源干细胞。
目的：探索不同成分培养基对人羊水间充质干细胞(human amniotic fluid-derived mesenchymal stem cells，h-AFMSCs)体外培养的影响，并分析最适培养基体外培养的 h-AFMSCs 的生物学特性，建立优化的h-AFMSCs培养方案。
方法：孕16~24周的孕妇产前检查中获得羊水进行原代及传代培养并采用6种不同的培养基成分对h-AFMSC进行培养。
结果与结论：从孕中期羊水中可分离出h-AFMSCs，体外使用低糖DMEM培养基(L-DMEM)+体积分数10%胎牛血清与合成培养基MESEN PRO体积比1∶1配制的培养基对其扩增促进作用最强。h-AFMSCs高表达CD29、CD73、CD90、CD105、CD166，低表达CD14、CD34、CD45及HLA-DR；分离培养的h-AFMSCs高表达的干细胞基因为OCT-4和Nanog；h-AFMSCs体外具有向成骨、成脂细胞分化的潜能且具有抑制淋巴细胞增殖的作用。提示孕中期羊水是低免疫原性的h-AFMSCs的良好细胞来源。
关键词：羊水；间充质干细胞；优化培养；生物学特性；培养基
doi:10.3969/j.issn.1673-8225.2012.10.016

关键词: 羊水, 间充质干细胞, 优化培养, 生物学特性, 培养基

Abstract:

BACKGROUND: At present, seed sources of stem cells engineering are diverse, and mesenchymal stem cells can be isolated from exfoliated cells in fetal amniotic fluid.
OBJECTIVE: To investigate the effects of the media with different elements on human amniotic fluid-derived mesenchymal stem cells (h-AFMSCs) cultured in vitro, and to analyze the biological characterization of h-AFMSCs at optimum medium cultured in vitro in order to optimize the culture strategy of h-AFMSCs.
METHODS: Amniotic fluid was extracted from pregnant women during 16 to 24 weeks of gestation, and then was used for primary culture and subculture. h-AFMSCs were cultured in media containing six different elements.
RESULTS AND CONCLUSION: h-AFMSCs were isolated from amniotic fluid in mid-trimester. The proliferative ability of h-AFMSCs was improved to a maximal extent in the mixed medium of low glucose Dulbecco's modified eagle medium containing 10% fetal bovine serum and MESEN PRO medium at a mixed ratio of 1:1. h-AFMSCs highly expressed CD29, CD73, CD90, CD105, CD166 and lowly expressed CD14, CD34, CD45, HLA-DR. Stem cells gene OCT-4 and Naong highly expressed in h-AFMSCs cultured in vitro. h-AFMSCs showed a higher osteogenic and adipogenic differentiation potential, and they could inhibit the proliferation of lymphocytes in vitro. It is indicated that mid-trimester amniotic fluid is a good cell source of h-AFMSCs with lower immunogenicity.

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