生理学报Acta Physiologica Sinica, April 25, 2003, 55(2): 153-159

Received 2002-07-01 Accepted 2002-12-03

This work was supported by the grants from 863 Program (#2002AA205051 and #2001AA216151) of Ministry of Science and Technology of China, from 973 Program (#2001CB509906) of Ministry of Science and Technology of China, and from Beijing Innovation Project (#A020220010190).

*Corresponding author. Tel: +86-10-66932203; Fax: +86-10-68167357; Email: peixt@nic.bmi.ac.cn

研究论文

人骨髓间充质干细胞在成年大鼠脑内的迁移及分化

侯玲玲1, 郑敏1, 王冬梅1, 袁红丰1, 李海民1, 陈琳1, 白慈贤1, 张涌2, 裴雪涛1,*

1军事医学科学院输血研究所、 军事医学科学院干细胞中心, 北京 100850;2 西北农林科技大学胚胎工程实验室,

陕西省杨凌 712100

摘要: 骨髓间充质干细胞(mesenchymal stem cells, MSCs)是目前倍受关注的一类具有多向分化潜能的组织干细胞, 体外可以分化为骨、 软骨、 脂肪等多种细胞。因此, MSCs是细胞治疗和基因治疗的种子细胞之一。为了探索MSCs的迁移和分化趋势, 为帕金森病(Parkinson disease, PD)的干细胞治疗提供理论和实验依据, 本实验将体外扩增并转染增强型绿色荧光蛋白(enhanced green fluorescent protein, EGFP)的人骨髓MSCs注入PD大鼠脑内纹状体, 观察了人骨髓MSCs在大鼠脑内的存活、 迁移、 分化以及注射MSCs前后大鼠的行为变化。结果表明, 人骨髓MSCs在大鼠脑内可存活较长时间(10周以上); 随着时间的延长, MSCs迁移范围扩大, 分布于纹状体、 胼胝体、 皮质以及脑内血管壁; 免疫组化法检测证实MSCs在大鼠脑内表达人神经丝蛋白(neurofilament, NF)、 神经元特异性烯醇化酶(neuron-specific enolase, NSE)以及胶质原纤维酸性蛋白(glial fibrillary acid protein, GFAP); PD大鼠的异常行为有所缓解, 转圈数由8.86±2.09 r/min下降到4.87±2.06 r/min, 统计学分析P<0.05为差异显著。以上观察结果表明, 骨髓MSCs有望成为治疗PD的种子细胞。

关键词: 骨髓间充质干细胞; 大鼠; 脑; 分化; 迁移

中图分类号: Q254

Migration and differentiation of human bone marrow mesenchymal stem cells in the rat brain

HOU Ling-Ling1, ZHENG Min1, WANG Dong-Mei1, YUAN Hong-Feng1, LI Hai-Min1, CHEN Lin1,BAI Ci-Xian1, ZHANG Yong2, PEI Xue-Tao1，*

1Beijing Institute of Transfusion Medicine, Beijing 100850; 2Northwest Sci-Tech University of Agriculture and Forestry, Yangling, Shanxi 712100

Abstract: Bone marrow mesenchymal stem cells (MSCs) are multipotent tissue stem cells that can be induced in vitro to differentiate into a variety of cells such as osteoblasts, chondrocytes and adipocytes. MSCs are useful vehicles for both cell and gene therapy for a variety of diseases. Here, we injected human MSCs with enhanced green fluorescent protein (EGFP) into the striatum of Parkinson disease (PD) rat and examined their survival, migration, differentiation, and the behavior changes in PD rats, which will provide a theoretical foundation and technical method for clinic PD therapy by stem cells. The results showed that human bone marrow MSCs can survive in rat brain for a long time (exceeding 70 d). MSCs were found in multiple areas of the rat brain including the striatum, the corpus callosum, contralateral cortex and even the brain vascular wall. Immunocytochemical staining suggested that implanted cells expressed human neurofilament (NF), neuron-specific enolase (NSE) and glial fibrillary acid protein (GFAP). At the same time, remission in abnormal behavior of the PD rats appeared. Rotation scores decreased gradually from 8.86±2.09 r/min pre-transplantation to 4.87±2.06 r/min 90 d post-transplantation (statistic result showed P<0.05).

Key words: mesenchymal stem cells; rat; brain; migration; differentiation