中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (49): 9235-.doi: 10.3969/j.issn.1673-8225.2010.49.026

• 干细胞基础实验 • 上一篇    下一篇

人羊膜间充质干细胞支持造血的体外实验

章守琴1，2，李维佳1，史明霞1，钱永海1   

  1. 1昆明医学院第一附属医院血液科，云南省昆明市650032；2 同济大学附属第十人民医院急诊科，上海市  200072
  • 出版日期:2010-12-03 发布日期:2010-12-03
  • 通讯作者: 史明霞，博士，副教授，昆明医学院第一附属医院血液科，云南省昆明市 650032 shmxia2002@sina.com
  • 作者简介:章守琴★，女，1976年生，安徽省合肥市人，汉族，昆明医学院在读硕士，主治医生，主要从事干细胞方面的研究。 zhshouqin@163.com
  • 基金资助:

    云南省应用基础研究面上项目(No.2007C136M)；昆明医学院第一附属医院院内博士启动金资助项目(No.2007bs05)。课题名称：人羊膜间充质干细胞支持造血和免疫学特性研究。

Supportive effect of human amniotic mesenchymal stem cells on hematopoiesis in vitro

Zhang Shou-qin 1,2, Li Wei-jia1, Shi Ming-xia1, Qian Yong-hai1   

  1. 1Department of Hematology, First Affiliated Hospital, Kunming Medical College, Kunming  650032, Yunnan Province, China;  2Department of Emergency, Tenth People’s Hospital Affiliated to Tongji University, Shanghai  200072, China
  • Online:2010-12-03 Published:2010-12-03
  • Contact: Shi Ming-xia, Doctor, Associate professor, Department of Hematology, First Affiliated Hospital, Kunming Medical College, Kunming 650032, Yunnan Province, China shmxia2002@sina.com
  • About author:Zhang Shou-qin★, Studying for master’s degree, Attending physician, Department of Hematology, First Affiliated Hospital, Kunming Medical College, Kunming 650032, Yunnan Province, China; Department of Emergency, Tenth People’s Hospital Affiliated to Tongji University, Shanghai 200072, China zhshouqin@163.com
  • Supported by:

    the General Program of Application Basic Research of Yunnan Province, No. 2007C136M*; the Doctor Priming Foundation Program of First Affiliated Hospital of Kunming Medical College, No. 2007bs05*

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摘要：

背景：骨髓是目前间充质干细胞的主要来源，但由于取材不便、细胞数量受年龄限制等原因，其应用具有一定局限性。近年来，羊膜作为间充质干细胞的新来源受到越来越广泛的关注。
目的：探讨人羊膜来源间充质干细胞在体外对造血干细胞的扩增是否有支持作用，以及怎样提高羊膜间充质干细胞和造血干细胞共移植成功率。
方法：利用组织块培养法，从足月分娩的人羊膜中分离培养羊膜间充质干细胞。密度梯度离心法分离脐血单个核细胞，采用免疫磁珠分选技术分选其中CD34+细胞。分别用脐血单个核细胞和CD34+细胞与羊膜间充质干细胞共培养，连续4周，每周计悬浮细胞浓度，并以骨髓间充质干细胞组及无滋养层组作为对照。采用集落形成实验，把扩增的脐血单个核细胞和CD34+细胞分别接种至甲基纤维素半固体培养基中，14 d后根据典型形态特征计数造血集落数。
结果与结论：羊膜间充质干细胞可促进人脐血单个核细胞和CD34+细胞扩增，扩增后两者在甲基纤维素半固体培养基中能够形成造血祖细胞集落，其造血支持作用与骨髓间充质干细胞相似，两者对比无明显统计学差异。提示，羊膜间充质干细胞体外具有与骨髓间充质干细胞相似的造血支持作用，有可能为造血干细胞体外扩增及临床间充质干细胞与造血干细胞联合移植、提高造血干细胞移植成功率提供一种更加理想的间充质干细胞新来源。

关键词: 羊膜, 间充质干细胞, 造血干细胞, 造血支持, 移植

Abstract:

BACKGROUND: Bone marrow (BM) is the main source of mesenchymal stem cell (MSCs) at present, but because of the inconvenience of isolation, and the age-related decrease in the number of MSCs, amnion as a new source of MSCs has been widespreadly concerned recently.
OBJECTIVE: To explore the supportive effects of MSCs derived from human amnions on hematopoiesis in vitro and to elevate the success rate of co-transplantation of amniotic mesenchymal stem cells (AMSCs) and hematopoietic stem cells.
METHODS: MSCs were isolated and cultured from human amnions after childbirth by the tissue pieces culture method. Mononuclear cells (MNCs) were isolated from umbilical cord blood using density gradient centrifugation. CD34+ cells were isolated by magnetic cell sorting. The MNC and CD34+ cells were co-cultured with AMSCs for four weeks respectively. The concentration of the floating cells was recorded weekly. And as the control, they were co-cultured with the BM-MSCs and deeder layer-free group respectively as well. The MNC and CD34-positive cells were plated in semisolid culture medium separately, and the colony formation was assessed at 14 days after the plating.
RESULTS AND CONCLUSION: AMSCs promoted the expansion of human cord blood MNCs and CD34+ cells, and also promoted the formation of hematopoietic progenitor cell colony in methylcellulose semisolid culture medium. Its capacity of hematopoitic support was similar to that of BM-MSCs, without significant difference. These indicate that in vitro AMSCs had the capacity for hematopoietic support similar to BM-MSCs. Thus, they may be an ideal source of stem cells for the expansion of hematopoietic stem cells in vitro and the co-transplantation of MSCs and hematopoietic stem cells to improve the success rate of clinical transplantations.

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