中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (19): 3555-3558.doi: 10.3969/j.issn.1673-8225.2011.19.032

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

人羊膜上皮细胞的干细胞特性

金  玲1，陈  剑2，吴  静3，徐锦堂3，周  清2，刘小勇2   

  1. 1深圳市宝安区人民医院，广东省深圳市518101
    2暨南大学附属第一医院眼科，广东省广州市  510630
    3暨南大学医学院眼科研究室，广东省广州市  510630
  • 收稿日期:2010-12-15 修回日期:2011-04-01 出版日期:2011-05-07 发布日期:2011-05-07
  • 通讯作者: 陈剑，博士，教授，主任医师，暨南大学附属第一医院眼科，广东省广州市510630 Drchenj@163.com
  • 作者简介:金玲☆，女，1976 年生，湖北省十堰市人，汉族，2009年暨南大学第一临床学院眼科学专业毕业，博士，主治医师，主要从事角膜病、角膜移植的临床和基础研究。 Jlingx2003@126.com
  • 基金资助:

    国家自然科学基金(30872808)，广东省重大科技计划基金项目(2003A3020104)，国务院侨办重点学科基金项目(51205004)资助。

Stem cell characteristics of human amniotic epithelial cells

Jin Ling1, Chen Jian2, Wu Jing3, Xu Jin-tang3, Zhou Qing2, Liu Xiao-yong2   

  1. 1People’s Hospital of Baoan District, Shenzhen  518101, Guangdong Province, China
    2Department of Ophthalmology, First Affiliated Hospital of Jinan University, Guangzhou  510630, Guangdong Province, China
    3Research Room of Ophthalmology, Medical College of Jinan University, Guangzhou  510630, Guangdong Province, China
  • Received:2010-12-15 Revised:2011-04-01 Online:2011-05-07 Published:2011-05-07
  • Contact: Chen Jian, Doctor, Professor, Chief physician, Department of Ophthalmology, First Affiliated Hospital of Jinan University, Guangzhou 510630, Guangdong Province, China Drchenj@163.com
  • About author:Jin Ling☆, Doctor, Attending physician, People’s Hospital of Baoan District, Shenzhen 518101, Guangdong Province, China Jlingx2003@126.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30872808*; Guangdong Key Science and Technology Planning Foundation, No. 2003A3020104*; Key Subject Foundation of Overseas Chinese Affairs Office of the State Council, No. 51205004 *

PDF

409

被引次数

17

摘要：

背景：研究发现人羊膜上皮细胞具有类似胚胎干细胞或多能干细胞的多向分化潜能，说明其可能是未来组织工程重建的一种新型种子细胞。
目的：研究体外培养的人羊膜上皮细胞的干细胞特性。
方法：取足月剖宫产的人羊膜组织，经酶消化法和差异黏附法获得纯度高的人羊膜上皮细胞，接种于含体积分数为10%胎牛血清的DMEM/F12培养基中进行原代和传代培养，用免疫荧光法和流式细胞仪检测法检测人羊膜上皮细胞表面胚胎干细胞的表面标记蛋白OCT-4和干细胞表面分子标记CD29、CD34、CD44、CD45、CD105的表达。
结果与结论：人羊膜上皮细胞在体外培养条件下呈上皮细胞特有的铺路石样外观，其胞浆中有OCT-4免疫荧光表达，其干细胞标记分子CD29、CD34的表达是阳性，但干细胞标记分子CD44、CD45、CD105的表达为阴性。结果表明人羊膜上皮细胞具有干细胞的某些特性，其可能是未来组织工程重建的一种新型种子细胞。

关键词: 人羊膜上皮细胞, 细胞培养, 胚胎干细胞, OCT-4, CD29, CD34

Abstract:

BACKGROUND: Studies have found that human amniotic epithelial cells (hAECs) have a multiple differentiation capacity similar as pluripotent stem cells or embryonic stem cells. It is indicated that hAECs may be a new kinds of seed cells in tissue engineering reconstruction in the further.
OBJECTIVE: To culture the hAECs in vitro and to investigate stem cell characteristics of cultured hAECs.
METHODS: The human amniotic tissue was delivered from full term caesarean, and the highly purified hAECs were obtained from enzyme digestion method and differently sticky method. The hAECs were seeded in DMEM/F12 medium obtaining 10% fetal bovine serum for the first time and serial subcultivation. The immunofluorescence and flow cytometry were used to detect the expression of OCT-4 of stem cell factor and Stem cell surface molecule marker CD29, CD34, CD44, CD45, CD105 in passaged hAECs.
RESULTS AND CONCLUSION: The cells in vitro looked like slabstone that epithelial cells specified. The results of immunofluorescence showed the expression of OCT-4 of stem cell factor in hAECs was positive. Detection of stem cell surface molecule marker by the flow cytometry showed the expression of CD29, CD34 on hAECs were positive, and the expression of CD44, CD45, CD105 on hAECs were negative. The findings show that the hAECs have some characteristics of embryonic stem cells, and may be a new kind of seed cells for the future tissue engineering reconstitution.

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