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$ ~" z# J" |9 w" }- Q* E- Q
(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4 }& w. C  w* _- A
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)& _) C1 T# |8 x4 P* J
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
! O1 p7 T# N* p% T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
4 Z9 z, o) P% H- A6 c9 D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2 F: h% v! c1 K5 c1 s2 f
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/ j6 P  i, K( X2 S8 x$ q
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+ N) ~8 F5 t" }: H! c
细胞特性。3 f, `/ v% x1 @# J
关键词:负压;骨髓间充质干细胞;成骨细胞;诱导
' A5 p) d$ ^# X6 Q) f: F* e中图分类号:R687 　　　文献标识码:A 　　　文章编号:167128259 (2008) 0520527203( b+ D  G2 j9 s# {) o. Q
Low2intensity interrupted negative pressure induces human marrow
9 X# K( u2 x( G7 i2 u" N. smesenchymal stem cells differentiating into osteoblasts
- x6 F0 J$ [" c  `$ `7 |Yang Zhi1 , Liu Miao1 , Zhang Yingang1 , Guo Xiong2 , Xu Peng3" r. t, h* Y- F( }- ]
(1. Department of Ort hopaedics , t he First Affiliated Hospital , Medical School of
* a% c8 E3 V' Z1 g! ?/ m1 HXipan J iaotong Univer sity , Xipan 710061 ; 2. Key Laboratory of Environment and Genes Related to" H: D2 D7 E6 ~  p9 |/ w, ^# A' y
Diseases of Minist ry of Education , Instit ute of Environmental and Endemic Diseases ,
/ e" }7 z8 r; }6 TXipan J iaotong Univer sity , Xipan 710061 ; 3. School of Material s Science and Engineering ,9 @/ E( ]; N0 V0 d! Y
Xipan J iaotong University , Xipan 710049 , China): C" A$ b! N( @4 `0 N' \
ABSTRACT: Objective 　To investigate t he ef f ect of low2intensity inter rup ted negative p ressure on t he; @) T/ E3 O( q* b
dif f e rentiation of human ma r row mesenchymal stem cells (MSCs) int o osteoblast in vit ro. Methods 　MSCs we re
6 y: H4 a" s, ^7 P3 o0 O: h3 B! Hisolated f rom adult ma r row using density gradient sep aration met hod , p assage 3 cells we re chosen t o induce
# S+ C9 F! E) s  D7 sdif f e rentiation ; t he dif f e rentiation of MSCs was examined by p hase2cont rast microscop y , t he dete rmination of
' S7 e, b$ x. k4 Qalkaline p hosp hatase (AL P) activities , alizarin2red staining and t he immunohist ochemist ry of type Ⅰcollagen and
/ R4 q! ^  H8 p5 C/ ^" bosteop rotege rin. Re sult s 　MSCs showed a typical app ea rance of osteoblasts af te r 2 weeksp induction by low2
1 _3 }" N6 H; c7 Ointensity inte r rup ted negative p ressure , t he activities of AL P we re increased signif icantly , calcium nodes we re seen
, j& ^4 v# r2 T% a7 [6 S$ nby alizarin2red staining , t he exp ressions of collagen type Ⅰ and osteop rotege rin were p ositive . Conclusion 　L ow2
( x# ^: P+ P0 N9 d: X" X; [intensity inte r rup ted negative p ressure could successf ully induce human MSCs int o osteoblasts and t he induced cells6 x) v6 q+ u! z
show cha racte ristics of osteoblasts .1 |% a: c' a% q( {, @9 y, D
KEY WORDS : negative p ressure ; ma r row mesenchymal stem cell ; osteoblast ; induction8 i  M; n0 u6 p5 v  @

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