中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (14): 2492-2496.doi: 10.3969/j.issn.1673-8225.2010.14.005

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

人脐带间充质干细胞分离培养及向脂肪与成骨细胞的分化

何绍清1，2，罗振宇1，2，刘秋英1，2，周向荣△2，3，邓铭权△2，3，罗  新4，姚润斯4，高  志5，王一飞○1，2   

  1. 1暨南大学生物医药研发基地，广东省广州市 510632；2广州(暨南)-香港细胞工程联合实验室，广东省广州市510632；3亚太干细胞科研中心有限公司，中国香港  999077；4暨南大学附属第一医院妇产科，广东省广州市 510632；   5克莱姆森大学生物工程系，美国南卡罗来纳州 29634-0905
  • 出版日期:2010-04-02 发布日期:2010-04-02
  • 通讯作者: 王一飞，教授，暨南大学生物医药基地，暨南大学生命工程与健康研究院，广东省广州市 510632
  • 作者简介:何绍清★，女，1984年生，江西省宜春市人，汉族，暨南大学在读硕士，主要从事间充质干细胞多能性的研究。 heshq106@163.com
  • 基金资助:

    广州(暨南)-香港细胞联合工程实验室干细胞分泌素抗衰老的研究与开发项目和国际科技合作项目干细胞技术在前叉韧带重建中的应用研究的资助。

Isolation of human umbilical cord mesenchymal stem cells and differentiation into adipocytes and osteblasts  

He Shao-qing 1, 2, Luo Zhen-yu 1, 2, Liu Qiu-ying 1, 2, Zhou Xiang-rong 2, 3, Deng Ming-quan 2, 3, Luo Xin4, Yao Run-si4, Gao Zhi5, Wang Yi-fei 1, 2   

  1. 1 Biomedicine Research and Development Centre of Jinan University, Guangzhou  510632, Guangdong Province, China; 2 Guangzhou (Jinan)-Hong Kong Joint Laboratory of Cell Engineering, Guangzhou  510632, Guangdong Province, China; 3 Asia-Pacific Stem Cell Research Centre Co., Ltd., Hong Kong  999077, China; 4 Department of Obstetrics and Gynecology, First Affiliated Hospital of Jinan University, Guangzhou  510632, Guangdong Province, China; 5 Department of Bioengineering, Clemson University, Clemson, SC 29634-0905, USA
  • Online:2010-04-02 Published:2010-04-02
  • Contact: Wang Yi-fei, Professor, Biomedicine Research and Development Centre of Jinan University, Guangzhou 510632, Guangdong Province, China; Guangzhou (Jinan)-Hong Kong Joint Laboratory of Cell Engineering, Guangzhou 510632, Guangdong Province, China
  • About author:He Shao-qing★, Studying for master’s degree, Biomedicine Research and Development Centre of Jinan University, Guangzhou 510632, Guangdong Province, China; Guangzhou (Jinan)-Hong Kong Joint Laboratory of Cell Engineering, Guangzhou 510632, Guangdong Province, China heshq106@163.com
  • Supported by:

    the Foundation of Guangzhou (Jinan)-Hong Kong Joint Laboratory*; International Cooperation Exchange for Science and Technology*

PDF

489

被引次数

21

摘要：

背景：研究报道显示人脐带间充质干细胞体外分离方法及效率、培养条件各不相同，并且尚未有统一的鉴定标准。因此建立高效、经济的培养体系十分必要。
目的：建立从人脐带组织分离培养间充质干细胞的方法，并进行向脂肪细胞和成骨细胞的诱导分化。
方法：采用组织块贴壁法和酶消化法分离纯化获得人脐带间充质干细胞，取对数生长期的第3代细胞，分析其细胞形态、增殖方式和某些免疫表型，并在体外诱导其向脂肪细胞和成骨细胞分化。
结果与结论：采用组织块贴壁法和酶消化法均能成功获得贴壁生长的人脐带间充质干细胞，并在体外扩增培养。流式细胞术分析结果显示，人脐带间充质干细胞强表达CD29，CD44，CD59，CD105，阴性表达CD28，CD31，CD34，CD45，CD40，CD86，HLA-DR。人脐带间充质干细胞在适当的诱导条件下能向脂肪和成骨细胞分化，体外能连续传代40次以上，且形态和表型保持稳定。这证实了人脐带组织存在间充质干细胞，且具有向脂肪和成骨细胞分化的潜能，因此将可能成为细胞治疗和组织工程新的种子细胞。

关键词: 脐带, 间充质干细胞, 脂肪细胞, 成骨细胞, 诱导分化

Abstract:

BACKGROUND: Culture condition, isolation method and efficiency are different in reported human umbilical cord-derived mesenchymal stem cells, which lack of unified identification standards. Therefore, it is necessary to establish a high-efficiency and economical culture system for human umbilical cord-derived mesenchymal stem cells (hUCMSCs).
OBJECTIVE: To isolate hUCMSCs and induced differentiate into adipocytes and osteblasts.
METHODS: The hUCMSCs were isolated form human umbilical cord by tissue adherence and digested with collagenase. The morphology, proliferation and immunophenotype of the 3rd passage cells were analyzed, and then cells were induced to osteogenic and adipogenic differentiation in vitro.
RESULTS AND CONCLUSION: The hUCMSCs isolated from human umbilical cord by tissue adherence and digested with collagenase could be cultured and proliferated in vitro. Flow cytometry analysis revealed that the hUCMSCs were positive for CD29, CD44, CD59, CD105, but were negative for CD40, CD86 and HLA-DR. These cells could be induced to differentiate into adipocytes and osteblasts under proper inducing conditions. The hUCMSCs retained the appearance and phenotype even after being expanded more than 40 passages in vitro. This confirmed that the existence of MSCs in human umbilical cord and they had the capacity of differentiating into adipocytes and osteblasts.

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