南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (05): 498-502.

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3H-TdR标记人骨髓间充质干细胞移植治疗mdx鼠的实验研究

柳太云1, 李经伦2, 姚晓黎1, 董群伟3, 苏全喜3, 冯善伟1, 李才明1, 曾缨1, 刘祖国1, 张成4, 刘长征1   

  1. 1. 中山大学第一医院神经内科, 广东, 广州, 510089;
    2. 中山大学基础医学院实验核医学教研室, 广东, 广州, 510089;
    3. 广州铁路局中心医院, 广东, 广州, 510080;
    4. 中山大学眼科中心, 广东, 广州, 510089
  • 出版日期:2005-05-20 发布日期:2005-05-20
  • 基金资助:
    收稿日期:2004-11-5。
    基金项目:卫生部临床学科重点项目(2001321);国家自然科学基金(30170337);博士点基金(20030558058);广东省自然科学基金(2003A3020102)资助;广东省重点基金(2003A3020102)
    作者简介:柳太云(1964- ),男,副主任医师,在读博士,现从事神经遗传性疾病研究,电话:020-87334192,E-mail:1tylxlf@gzsums.edu.cn
    通讯作者:张成,电话:020-86332387;E-mail:czyrn@gzsums.edu.cn

Transplantation of 3H-thymidine-labeled human bone marrow-derived mesenchymal stem cells in mdx mice

LIU Tai-yun1, LI Jing-lun2, YAO Xiao-li1, DONG Qun-wei3, SU Quan-xi3, FENG Shan-wei1, LI Cai-ming1, ZENG Ying1, LIU Zu-guo1, ZHANG Cheng4, LIU Chang-zheng1   

  1. 1. 中山大学第一医院神经内科, 广东, 广州, 510089;
    2. 中山大学基础医学院实验核医学教研室, 广东, 广州, 510089;
    3. 广州铁路局中心医院, 广东, 广州, 510080;
    4. 中山大学眼科中心, 广东, 广州, 510089
  • Online:2005-05-20 Published:2005-05-20

摘要: 目的 探讨人骨髓间充质干细胞移植mdx鼠能否修复骨骼肌肌膜病变及干细胞在其体内的分布特点.方法 用氚-脱氧胸腺嘧啶(3H-TdR)标记人骨髓间充质干细胞(hBM-MSCs),以每只鼠干细胞1×107/0.3 ml经尾静脉注入18只免疫抑制的mdx鼠(8~10周龄).于移植后24 h、48 h、2周、1月、2月、4月处死鼠,分别测定各组织器官的放射活性;用免疫荧光法检测骨骼肌肌膜营养障碍基因(dystrophin,Dys)表达情况.结果 移植后1月,多数组织、器官放射活性显著高于对照组,其中以骨髓、肝、脾放射活性增高更为明显.大多数组织器官放射活性随时间推移逐渐下降,而骨骼肌放射活性的从2周时开始逐渐升高,于1月时达到高峰(27.6±3.3 Bq/mg湿重).1月之后,骨髓、骨骼肌放射活性仍保持较高水平.免疫荧光显示移植后24 h、48 h、2周骨骼肌Dys免疫荧光呈阴性,1月后呈阳性,1月、2月、4月阳性率分别为:6.6%、8.4%、8.9%.结论 hBM-MSCs能植入mdx鼠体内,并融合、分化为骨骼肌,部分修复骨骼肌肌膜病变;hBM-MSCs移植入mdx鼠后早期主要分布在骨髓、肝、脾,后期主要分布在骨髓、骨骼肌.

Abstract: Objective To investigate the feasibility of using human bone marrow-derived mesenchymal stem cells (hBM- MSCs) for repairing the skelatal muscle sarcolemma lesions in mdx mice and characterize the distribution of the transplanted hBM-MSCs.Methods Eighteen 8-to 10-week-old immunosuppressed mdx mice received transplantation with 1×107 of hBM-MSCs (the fifth passage) with 3H-thymidine (3H-TdR) labeling by injection of the cells into the tail vein. The mice were killed at 24 h, 48 h, 2 weeks, and 1, 2 and 4 months after the transplantation, respectively, to measure the radioactivity in the tissues and organs. Dystrophin expression on the sarcolemma was detected by immunofluorescence analysis.Results One month after transplantation, the mice with cell transplantation showed greater radioactivity in most of the tissues and organs than the control mice, especially in the bone marrow, liver and spleen. The radioactivity was then gradually lowered but in the skeletal muscle, the radioactivity increased progressively since 2 weeks after transplantation, reaching the peak of 27.65±3.53 Bq/mg at 1 month. Compared with that in the control mice, the radioactivity in the bone marrow and skeletal muscle was persistently higher in mice with cell transplantation 1 month after transplantation. No dystrophin-positive cells were found in the mdx mice at 2 weeks but detected at 1 month. The percentage of dystrophin-positive fibers in each section ranged from 6.6% (1 month) to 8.9% (4 months). Conclusions hBM-MSCs engrafted in immunosuppressed mdx mice may differentiate into skeletal muscle cells to repair the pathological lesion of the skeletal muscle sarcolemma. The hBM-MSCs reside mainly in the bone marrow, liver and spleen in the early stage following transplantation, homing into the bone marrow and skeletal muscle later.

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