杨 治1 ,刘 淼1 ,张银刚1 ,郭 雄2 ,许 鹏39 A0 b: U+ Q4 r V2 `9 a
(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$ k5 `6 S( Z& l) A4 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) l& L1 U1 ?; e0 F
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, ]6 @4 j K" R% j- O6 y5 V
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! m- ~' V' _# v
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% o# S# W/ q, A; y6 @: n
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% o8 Z1 h- S' T
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9 q$ x1 s, G! X8 g m& ]
细胞特性。4 V: H& ?9 Z9 P( R8 H
关键词:负压;骨髓间充质干细胞;成骨细胞;诱导7 J8 R4 H% I& t
中图分类号:R687 文献标识码:A 文章编号:167128259 (2008) 05205272038 W" x! }6 G& Q7 f4 B
Low2intensity interrupted negative pressure induces human marrow ^0 \6 i8 r4 m3 u
mesenchymal stem cells differentiating into osteoblasts4 r. F0 |0 y" \9 F. T
Yang Zhi1 , Liu Miao1 , Zhang Yingang1 , Guo Xiong2 , Xu Peng3 " S4 {% z% ^7 [* V(1. Department of Ort hopaedics , t he First Affiliated Hospital , Medical School of% k. v6 b, S2 o7 Z
Xipan J iaotong Univer sity , Xipan 710061 ; 2. Key Laboratory of Environment and Genes Related to$ A: i' D C% T& S% b
Diseases of Minist ry of Education , Instit ute of Environmental and Endemic Diseases ,/ n1 t* r+ w$ ^9 O, _# K
Xipan J iaotong Univer sity , Xipan 710061 ; 3. School of Material s Science and Engineering , + |2 m9 ]0 }0 R6 R3 O% r7 ?. ~Xipan J iaotong University , Xipan 710049 , China) 5 X& d" H& ?1 P. c, V2 _6 TABSTRACT: Objective To investigate t he ef f ect of low2intensity inter rup ted negative p ressure on t he) W+ a6 m ^0 S3 k* p
dif f e rentiation of human ma r row mesenchymal stem cells (MSCs) int o osteoblast in vit ro. Methods MSCs we re u# H( S" n' l
isolated f rom adult ma r row using density gradient sep aration met hod , p assage 3 cells we re chosen t o induce. O7 H# }* u5 h5 b3 o6 Z
dif f e rentiation ; t he dif f e rentiation of MSCs was examined by p hase2cont rast microscop y , t he dete rmination of8 S8 T9 n7 U, x2 t- Q0 R5 p' q
alkaline p hosp hatase (AL P) activities , alizarin2red staining and t he immunohist ochemist ry of type Ⅰcollagen and : ~ j7 y3 X, l( P$ c2 `% i3 Q Oosteop rotege rin. Re sult s MSCs showed a typical app ea rance of osteoblasts af te r 2 weeksp induction by low2 {6 T( g6 ^. B9 wintensity inte r rup ted negative p ressure , t he activities of AL P we re increased signif icantly , calcium nodes we re seen - j1 d* V3 ], x7 B/ ]by alizarin2red staining , t he exp ressions of collagen type Ⅰ and osteop rotege rin were p ositive . Conclusion L ow2 7 O( W4 M" ^/ s mintensity inte r rup ted negative p ressure could successf ully induce human MSCs int o osteoblasts and t he induced cells 2 X) ?- `7 o3 N) A8 H$ |show cha racte ristics of osteoblasts . 6 o, W1 b2 o# b& @& i) WKEY WORDS : negative p ressure ; ma r row mesenchymal stem cell ; osteoblast ; induction \/ g U: m r3 l! t1 c6 V: G$ _ `( E, z; Z9 g- ~/ s0 A* W
附件为全文，请大家支持一下 回复本贴就可以下载了 + K7 m' l! ~* E: w( Z$ [, \/ @; i4 \$ U" R 作者: xuliangytu 时间: 2009-12-7 20:56