人骨髓间充质干细胞在成年大鼠脑内的迁移及分化

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/ W1 r3 V( H& D1 w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 ,
. k' T! y+ E8 x/ Y5 N" l5 W裴雪涛1 , 3* B  E/ J  I' s$ R9 J
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,
3 L; s" _4 Y# {, }/ a" \$ b陕西省杨凌7121000 U) Y; Y6 l; I2 |. @/ Y
摘　要: 　骨髓间充质干细胞(mesenchymal stem cells , MSCs) 是目前备受关注的一类具有多向分化潜能的组织干细胞,
2 ?* d4 {% e1 q5 t. v' ]6 {0 t( X( e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
4 E6 R" [+ Q+ O1 ^! e( n+ ?& u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6 A' F; i! E" X6 t( z
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
- [# R! u8 k, J  p! Y3 }5 b, M( kMSCs 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
  S1 j5 W! f; L0 e0 X9 G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- U2 s8 _& ~2 A2 Y$ q4 h
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(neuron2specific eno2
+ t- U  e$ S& D( I! }6 @lase , 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8186 ±* J6 U6 y! q5 w% z4 H0 I/ b8 R
2109 r/ min 下降到4187 ±2106 r/ min , 统计学分析P < 0105 为差异显著。以上观察结果表明, 骨髓MSCs 有望成为治疗  d2 H& g+ J( v( y2 n' s
PD 的种子细胞。
7 G5 l# F* D+ b) ^关键词: 骨髓间充质干细胞; 大鼠; 脑; 分化; 迁移
  E/ k$ n9 X6 G6 i' S中图分类号: Q254
( K: P( P+ t" l! K" f' D4 qMigration and differentiation of human bone marrow mesenchymal stem9 j* q" t4 B: g; x5 z
cells in the rat brain7 U: D+ j1 G7 _3 \% z3 F9 a' K
HOU Ling2Ling1 , ZHENGMin1 , WANG Dong2Mei1 , YUAN Hong2Feng1 , LI Hai2Min1 , CHEN Lin1 ,
$ l$ p. A; }, m, \1 p8 n! rBAI Ci2Xian1 , ZHAN G Yong2 , PEI Xue2Tao1 , 3
4 a5 A1 d: u1 c% W* T1Beijing Instit ute of Transf usion Medicine , Beijing 100850 ; 2 Northwest Sci2Tech University of A gri2
$ f6 E" W2 D# Ycul t ure and Forest ry , Yangling , S hanxi 712100
1 J" p: W# a. L( q( iAbstract : 　Bone marrow mesenchymal stem cells (MSCs) are multipotent tissue stem cells that can be induced' x2 B! X% J  t
in vit ro to differentiate into a variety of cells such as osteoblasts , chondrocytes and adipocytes. MSCs are useful" E8 W" y1 g  ?# Z: H' m

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vehicles for both cell and gene therapy for a variety of diseases. Here , we injected human MSCs with enhanced
3 R' }/ T1 i" W2 M; v3 z, cgreen fluorescent protein (EGFP) into the striatum of Parkinson disease (PD) rat and examined their survival ,
1 S. B9 }& ~- \! @! N! wmigration , differentiation , and the behavior changes in PD rats , which will provide a theoretical foundation and) ^7 |  z' h) U" p# t
technical method for clinic PD therapy by stem cells. The results showed that human bone marrow MSCs can/ n- q0 L5 s. M& v- W: M
survive in rat brain for a long time (exceeding 70 d) . MSCs were found in multiple areas of the rat brain includ2
; l$ N# g8 h' q/ m' J1 t* D7 Eing the striatum , the corpus callosum , contralateral cortex and even the brain vascular wall. Immunocytochemical, s3 u  A+ {6 [/ w/ F% I
staining suggested that implanted cells expressed human neurofilament (NF) , neuron2specific enolase (NSE) and; v) I% h0 V1 B3 p. ^* Z7 E4 E
glial fibrillary acid protein (GFAP) . At the same time , remission in abnormal behavior of the PD rats appeared.* \% @3 u; W" ^- ]; \  `& W+ j
Rotation scores decreased gradually from 8186 ±2109 r/ min pre2transplantation to 4187 ±2106 r/ min 90 d post27 [: C% v' |, u$ W
transplantation (statistic result showed P < 0105)
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