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
) `* d% m8 I2 F+ {(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
  A  ]' |4 J- K, Z9 G0 t. B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)" O+ w6 R' }, ]2 V8 W1 }8 [! S5 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
- @; k# N+ h  J0 _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. i3 a6 e" }& @- d' B: E
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' d/ w3 f) y- s; G7 Y
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# w7 H& l( C6 z4 Y' ~, S8 Z
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* t, x6 Y1 B& I% I* |' }2 R& G
细胞特性。9 r% p6 c/ Y" F& e
关键词:负压;骨髓间充质干细胞;成骨细胞;诱导3 b* o9 X" _- ?" ]% _: {+ L* \
中图分类号:R687 　　　文献标识码:A 　　　文章编号:167128259 (2008) 0520527203) c; D& o, v. v5 L
Low2intensity interrupted negative pressure induces human marrow
( u, X2 W5 Q: b9 }8 jmesenchymal stem cells differentiating into osteoblasts
9 ?% m5 a* J" E3 v$ w. yYang Zhi1 , Liu Miao1 , Zhang Yingang1 , Guo Xiong2 , Xu Peng3
" `: w" c" w. C. |: L( F* c(1. Department of Ort hopaedics , t he First Affiliated Hospital , Medical School of! [( i) r3 ?3 S
Xipan J iaotong Univer sity , Xipan 710061 ; 2. Key Laboratory of Environment and Genes Related to6 j  G3 k! I5 H: h4 k
Diseases of Minist ry of Education , Instit ute of Environmental and Endemic Diseases ,: r- @* I/ E) q
Xipan J iaotong Univer sity , Xipan 710061 ; 3. School of Material s Science and Engineering ,
' M" P+ x/ @- s: d5 z. S5 R7 EXipan J iaotong University , Xipan 710049 , China)2 y# d2 }7 M$ L0 q9 J) @' g
ABSTRACT: Objective 　To investigate t he ef f ect of low2intensity inter rup ted negative p ressure on t he
: K6 ^. a2 o1 J( g! Bdif f e rentiation of human ma r row mesenchymal stem cells (MSCs) int o osteoblast in vit ro. Methods 　MSCs we re
1 t! G3 m2 m6 }( w/ b2 N* _isolated f rom adult ma r row using density gradient sep aration met hod , p assage 3 cells we re chosen t o induce
) @# V; U4 A6 C3 N( j8 Ydif f e rentiation ; t he dif f e rentiation of MSCs was examined by p hase2cont rast microscop y , t he dete rmination of" `# b& c4 T, ^+ L) s8 m: k! l
alkaline p hosp hatase (AL P) activities , alizarin2red staining and t he immunohist ochemist ry of type Ⅰcollagen and/ N5 _9 e4 Z" O  m
osteop rotege rin. Re sult s 　MSCs showed a typical app ea rance of osteoblasts af te r 2 weeksp induction by low2: o0 Y8 L9 {- o% i; b3 z( _8 K
intensity inte r rup ted negative p ressure , t he activities of AL P we re increased signif icantly , calcium nodes we re seen6 t, D& e3 q! a% o
by alizarin2red staining , t he exp ressions of collagen type Ⅰ and osteop rotege rin were p ositive . Conclusion 　L ow29 H3 V) _0 X+ b
intensity inte r rup ted negative p ressure could successf ully induce human MSCs int o osteoblasts and t he induced cells' Q  I* u2 P8 b6 }6 [; `
show cha racte ristics of osteoblasts .+ x! m% L: g6 i( z: |+ m7 ~# w
KEY WORDS : negative p ressure ; ma r row mesenchymal stem cell ; osteoblast ; induction
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yuweiliang

好啊谢谢诶

xuliangytu

多谢