Migration of intravenously grafted mesenchymal stem cells to injured

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* a* i$ I- p7 g" s) O1 ^Migration of intravenously grafted mesenchymal stem cells to injured
, l# E0 U( v! ]2 Z; aheart in rats
) k! r# n9 G9 I9 i9 BJIANG Wen-Hui, MA Ai-Qun*, ZHANG Yan-Min, HAN Ke, LIU Yu, ZHANG Zeng-Tie, WANG Ting-Zhong,
3 {& ]1 K* w9 K5 g3 tHUANG Xin, ZHENG Xiao-Pu
5 O- o7 v9 c# r9 x+ V! m7 O3 }Cardiovascular Department, the First Hospital of Xi’an Jiaotong University; Key Laboratory of Environment and Genes Related to$ F3 k' A- r; T5 Y! @
Diseases (Xi’an Jiaotong University), Ministry of Education, Xi’an 710061, China! D- M' Z$ _0 a2 r* C
Abstract: The present study aimed to determine the role of tissue injury in migration of mesenchymal stem cells (MSCs) intravenously. Q7 W+ k2 Q; c9 t. ?
transplanted into heart and to establish experimental basis for improving stem cell therapy in its targeting and effectiveness. MSCs were4 s$ v3 c# f1 \- ~% y# \, w
isolated from bone marrow of male Sprague-Dawley rats and purified by density centrifuge and adhered to the culture plate in vitro.! ?; e/ z, a  n+ N4 G
Female rats were divided randomly into four groups. Myocardial ischemia (MI) transplanted group received MSCs infusion through2 N  U+ Q( p# \5 P9 N4 K3 R' N
tail vein 3 h after MI and compared with sham-operated group or normal group with MSCs infusion, or control group received culture" f4 E0 E- F5 H; g
medium infusion. MI was created in female rats by ligating the left anterior descending coronary artery. The heart was harvested 1 week9 b- B' V! ?  x  }
and 8 weeks after transplantation. The characteristics of migration of MSCs to heart were detected with expression of sry gene of Y- h" f# t8 T1 e- K, Y
chromosome by using fluorescence in situ hybridization (FISH). Ultrastructural changes of the ischemic myocardium of the recipient
6 W6 S' O- N! L8 [3 `0 U1 \. v" krats were observed by transmission electron microscope (TEM). One week or 8 weeks after transplantation, sry positive cells were
/ q& g3 ]) X! O% Z5 nobserved in the cardiac tissue in both of MI transplanted group and sham-operated group, the number of sry positive cells being) x  o0 Y% L/ R8 [" Y
significantly higher in MI transplanted group (P<0.01). No significant difference was found in the number of sry positive cells between8 {8 S) \  M, ]# A% R
1 week and 8 weeks after transplantation. No sry positive cells were observed in the hearts of control and normal group. In addition, the
: h9 r* g0 r" J9 dultrastructure of some cells located in the peri-infarct area of MI rats with MSCs transplantation was similar to that of MSCs cultured
; ^7 Z) {* F0 Z6 [in vitro. These results indicate that MSCs are capable of migrating towards ischemic myocardium in vivo and the fastigium of migration  B- g  E! ^! }2 F/ v+ b
might appear around 1 week after MI. The tissue injury and its degree play an important role in the migration of MSCs.
2 _$ Z7 w8 ]$ q* ^2 U$ D* u( w8 ?9 gKey words: mesenchymal stem cell; tissue injury; migration; heart8 _0 d5 G# A* s6 \( e
骨髓间质干细胞向大鼠损伤心肌组织的迁移
5 z( i, j$ H) k蒋文慧，马爱群*，张艳敏，韩克，刘昱，张增铁，王亭忠，黄 欣，郑小璞
# N' x8 j9 r2 |西安交通大学第一医院心血管内科, 教育部环境与疾病相关基因重点实验室，西安710061
/ x3 e. z9 J1 |  D6 x摘 要：实验旨在动态观察骨髓间充质干细胞(mesenchymal stem cells, MSCs)向不同微环境下心肌组织的迁移特点，明确组织2 G- [% h7 j( A7 A/ ]2 q* L( d1 @
损伤在干细胞迁移中的作用，为提高干细胞治疗的靶向性和高效性奠定初步试验基础。分离纯化雄性Sprague-Dawley (SD)大
$ M- _# T( r* ~; Z2 c鼠的骨髓MSCs，输注入雌性SD 大鼠。实验分为4 组：正常大鼠+MSCs 移植组，假手术+MSCs 移植组，心肌缺血+MSCs
5 h' L$ q) X9 z- B移植组，心肌缺血对照组(心肌缺血+ 培养基移植)。结扎冠状动脉前降支制造心肌缺血模型，将相等数量的雄性MSCs 经尾* Z, }  P' N0 a& [; N2 Z
静脉注射移植入前3 组雌性大鼠体内，对照组注射等体积培养基，分别于移植后1 周及8 周取心脏组织标本，采用荧光原位1 y( i$ s: \3 E7 _0 O6 n2 ?( n% m
杂交方法(fluorescence in situ hybridization, FISH)检测大鼠Y 染色体雄性鉴别基因sry 片段的表达，用透射电镜观察大鼠心肌组( m+ U. \, U& t$ F' [8 w
织超微结构改变。结果发现，移植后1 周和8 周，正常大鼠移植组和对照组大鼠的心肌组织中均未见sry 基因的表达，但假% u1 g5 ^( x. D5 Y& m# }* I" u
手术移植组和心肌缺血移植组的心肌组织中均可见sry 基因的表达，心肌缺血移植组的Y 染色体sry 基因阳性细胞数量在两个7 ^  a8 I7 _! V1 S9 M' j- G2 T; L
时间点均显著高于假手术移植组(P<0.01)。分别比较心肌缺血移植组和假手术组在移植后1 周和8 周的Y 染色体sry 基因阳性细
* u- G' J6 _) [% C7 p4 j胞的数量，两个时间点无明显差异。心肌组织的超微结构观察发现心肌缺血移植组大鼠的心肌梗死周边区域可见一些细胞，: o, H% [# U2 T' X8 o; P
其形态类似于体外培养的MSCs。研究结果提示MSCs 具有向损伤心肌组织迁移的特性，迁移的高峰期可能在组织损伤1 周左右，组织损伤及其程度在干细胞迁移中起重要作用。) s+ I( x1 `+ A- P5 m$ z8 ~, r
关键词：骨髓间质干细胞；组织损伤；迁移；心脏
7 B* m! |$ Y- V  I中图分类号：R364.3; R542.2
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