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+ ]- |7 S5 }8 _4 O
(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
2 ?3 J  _& \, H' l9 z4 h$ A0 r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)
  {  Z# Z8 D3 \6 t7 [! x' R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$ Q# Q& j" r7 w8 k* ^
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 ?2 g, N. s# u" w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) c) I" T* v; P5 B- r
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
: U% [! C/ P# {9 n3 m% ]/ \/ v1 D+ @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/ ^4 w% O$ ^) X9 Z
细胞特性。
1 T  k; v  D! E; i) l& H: G关键词:负压;骨髓间充质干细胞;成骨细胞;诱导+ s9 G) R8 P  D3 q' p! W
中图分类号:R687 　　　文献标识码:A 　　　文章编号:167128259 (2008) 0520527203$ o5 u( m3 Z3 }8 n0 k
Low2intensity interrupted negative pressure induces human marrow8 V2 N! Y2 o& a8 b* z2 X
mesenchymal stem cells differentiating into osteoblasts4 p$ `! [  x; t5 u9 _7 ]& ^
Yang Zhi1 , Liu Miao1 , Zhang Yingang1 , Guo Xiong2 , Xu Peng3; N, A4 Z4 _. Q5 m" t) o: H
(1. Department of Ort hopaedics , t he First Affiliated Hospital , Medical School of
0 z" p" |% g6 n1 JXipan J iaotong Univer sity , Xipan 710061 ; 2. Key Laboratory of Environment and Genes Related to8 L, @  w  J5 n$ A4 {; D7 d4 d
Diseases of Minist ry of Education , Instit ute of Environmental and Endemic Diseases ,
' ?! S; h! d' B  T3 F1 _6 DXipan J iaotong Univer sity , Xipan 710061 ; 3. School of Material s Science and Engineering ,  i8 S1 [, W6 U4 D# [
Xipan J iaotong University , Xipan 710049 , China)! s/ N, j% R* ]1 z
ABSTRACT: Objective 　To investigate t he ef f ect of low2intensity inter rup ted negative p ressure on t he
; @$ ?* @% z; U  l& N2 X9 Bdif f e rentiation of human ma r row mesenchymal stem cells (MSCs) int o osteoblast in vit ro. Methods 　MSCs we re
8 w( E! b, K9 G# c, v/ u1 \isolated f rom adult ma r row using density gradient sep aration met hod , p assage 3 cells we re chosen t o induce3 X3 }- Q& r- i
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
. S7 G$ }! x2 ~$ e9 D0 `alkaline p hosp hatase (AL P) activities , alizarin2red staining and t he immunohist ochemist ry of type Ⅰcollagen and* u" m! |' Y: D* |7 z4 K
osteop rotege rin. Re sult s 　MSCs showed a typical app ea rance of osteoblasts af te r 2 weeksp induction by low29 t& Q7 U2 k0 ~6 G8 g4 v" |1 M9 t& Z
intensity inte r rup ted negative p ressure , t he activities of AL P we re increased signif icantly , calcium nodes we re seen- s( O# h9 ]! T$ Y6 i
by alizarin2red staining , t he exp ressions of collagen type Ⅰ and osteop rotege rin were p ositive . Conclusion 　L ow2
8 _. c; g% I# s# kintensity inte r rup ted negative p ressure could successf ully induce human MSCs int o osteoblasts and t he induced cells; R7 n1 N& P& w& G8 c$ s! s
show cha racte ristics of osteoblasts .) o) g9 e7 i% I6 {5 A7 l& s
KEY WORDS : negative p ressure ; ma r row mesenchymal stem cell ; osteoblast ; induction
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xuliangytu

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yuweiliang

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