低强度间断负压诱导人骨髓间充质干细胞向成骨细胞的分化

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杨　治1 ,刘　淼1 ,张银刚1 ,郭　雄2 ,许　鹏3
0 i. Q  [0 w  n9 T(1. 西安交通大学医学院第一附属医院骨科,陕西西安　710061 ;2. 西安交通大学环境与疾病相关基因
; m. r; f. y5 L, V" c# C! m$ W教育部重点实验室,环境与地方病研究室,陕西西安　710061 ;3. 西安交通大学材料学院,陕西西安　710049)6 a3 {- c) [6 j1 r! x7 [$ Q1 U4 p
摘要:目的　利用低强度间断负压诱导人骨髓间充质干细胞(MSCs) 向成骨细胞定向分化。方法　分离人骨髓,利用! E) B. V5 E  h9 ?6 Q! Z
梯度离心法进行MSCs 的培养,取第三代细胞利用低强度间断负压诱导分化;倒置显微镜下观察细胞的形态,检测碱" ?0 R5 o  q1 B0 f$ a/ Y
性磷酸酶(AL P) 活性,茜素红染色观察钙结节形成,免疫组织化学检测Ⅰ型胶原和骨保护素的表达。结果　低强度间
( G4 H2 r8 n! |& g断负压诱导2 周后,细胞呈现出成骨细胞形态,AL P 活性明显增强,茜素红染色可见钙结节形成, Ⅰ型胶原和骨保护
" l: P; M% L+ l0 w素表达阳性。结论　利用低强度间断负压可以成功的诱导MSCs 向成骨细胞定向分化,诱导的细胞具有典型的成骨
; _, m3 }. w7 ?0 a  s5 i. F* [细胞特性。
8 T) s( o8 r. i; J$ C: j: F关键词:负压;骨髓间充质干细胞;成骨细胞;诱导; a  ?3 s# I3 `/ b+ m! W
中图分类号:R687 　　　文献标识码:A 　　　文章编号:167128259 (2008) 0520527203
. F4 ]* o1 F+ w# v5 HLow2intensity interrupted negative pressure induces human marrow8 c# l8 x+ C6 @1 h3 d9 @& t+ X, e
mesenchymal stem cells differentiating into osteoblasts
( a, f% {7 r7 SYang Zhi1 , Liu Miao1 , Zhang Yingang1 , Guo Xiong2 , Xu Peng3
! O5 v/ n5 ?0 r- ^(1. Department of Ort hopaedics , t he First Affiliated Hospital , Medical School of
6 |/ z. ?( w) ?Xipan J iaotong Univer sity , Xipan 710061 ; 2. Key Laboratory of Environment and Genes Related to
5 _( J3 H4 }7 v$ i- M0 VDiseases of Minist ry of Education , Instit ute of Environmental and Endemic Diseases ,
: V" R" {: @" D2 v- k3 B7 sXipan J iaotong Univer sity , Xipan 710061 ; 3. School of Material s Science and Engineering ,3 z# a6 v  n+ C; K) k: k- i& S
Xipan J iaotong University , Xipan 710049 , China)3 [) B$ n1 u8 Y+ Z1 M' Q$ o& ~: F
ABSTRACT: Objective 　To investigate t he ef f ect of low2intensity inter rup ted negative p ressure on t he* N0 ~" X; M3 u( V5 F" \
dif f e rentiation of human ma r row mesenchymal stem cells (MSCs) int o osteoblast in vit ro. Methods 　MSCs we re
! N$ I7 b) u4 Z' N) p* E, m' x' @isolated f rom adult ma r row using density gradient sep aration met hod , p assage 3 cells we re chosen t o induce4 f3 F! H# Z0 o  ?9 o
dif f e rentiation ; t he dif f e rentiation of MSCs was examined by p hase2cont rast microscop y , t he dete rmination of$ I% s, @# R! q; o2 F( E% ?7 l4 D
alkaline p hosp hatase (AL P) activities , alizarin2red staining and t he immunohist ochemist ry of type Ⅰcollagen and
$ G% [0 G5 \: i* g2 R8 x* Mosteop rotege rin. Re sult s 　MSCs showed a typical app ea rance of osteoblasts af te r 2 weeksp induction by low2
/ r; f8 {- ~- Gintensity inte r rup ted negative p ressure , t he activities of AL P we re increased signif icantly , calcium nodes we re seen
4 I+ r" _$ q; uby alizarin2red staining , t he exp ressions of collagen type Ⅰ and osteop rotege rin were p ositive . Conclusion 　L ow2
) l3 F% x+ O) z! {5 k# B$ x6 Fintensity inte r rup ted negative p ressure could successf ully induce human MSCs int o osteoblasts and t he induced cells
; q+ z  J1 N; }# y  c* Zshow cha racte ristics of osteoblasts ./ y9 B9 m) M) s* a" l
KEY WORDS : negative p ressure ; ma r row mesenchymal stem cell ; osteoblast ; induction
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