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本帖最后由 细胞海洋 于 2010-5-17 23:06 编辑 8 N1 }9 ?6 s+ Y/ x5 s9 u- V
9 m- i3 J2 @7 J- G" p* G安方梅1 , 邓永东2 , 赵正斌2 ,陈 红2
J2 V ]- e2 w! v(1. 兰州大学临床医学院,甘肃兰州730000 ;2. 兰州大学第一医院感染科,甘肃兰州730000)
, o; y7 C( A) \8 ]摘 要:目的 研究肝硬化患者骨髓间充质干细胞(MSCs) 的体外扩增方法及其生长特性和特异性表面标记,为肝
' }6 h& N- S( _9 D0 [组织工程“种子细胞”的选择提供新的理论依据,为进一步研究自体骨髓间充质干细胞移植治疗肝衰竭时移植细胞, U* `# a* b! I& c
在体内的调控机制打下基础。方法 无菌抽取4 例肝硬化志愿者骨髓3～4 mL ,年龄30～40 岁,利用密度梯度离
t9 P+ t/ d) @& [, i3 }1 E心法联合贴壁筛选法逐步筛选MSCs ,获取MSCs ,倒置显微镜下逐日观察细胞生长特性,细胞计数,绘制细胞生长
8 ~( t) T* k+ P: y- Y曲线,流式细胞仪检测细胞表面标志CD44 、CD45 。结果 原代培养细胞生长潜伏期0～9 d ,对数增殖期为10～19
( s) W" `% X; _5 y8 Qd ,生长平台期为20～24 d ;传代细胞生长周期较原代细胞快,细胞生长潜伏期为4～24 h ,对数增殖期为3～12 d , [9 \0 g, a4 L- @
13～15 d 进入生长平台期。肝硬化患者骨髓MSCs 高表达CD44 ,低表达CD45 。结论 肝硬化患者骨髓MSCs 体+ \" o9 S6 j/ I! M' O
外生长良好,性质稳定,可以作为“种子细胞”用于自体骨髓干细胞移植和生物人工肝系统。
1 G# q" Q" T4 |- C6 \4 s$ _关键词:肝硬化;骨髓间充质干细胞; 自体骨髓干细胞;移植
) `! c( w* z: g1 E/ i E7 c中图分类号:R33 文献标识码:A
0 r) m z, ~6 X0 EI n vit ro culture and growth characteristics of bone marrow
& v* \3 `5 y, q! h5 I' nmesenchymal stemcells from liver cirrhosis patients$ D& e9 L: A: u! s# M, y I
AN Fang2mei1 ,DEN G Yong2dong2 , ZHAO Zheng2bin2 , CHEN Hong2% l- x8 ]" B7 a! `5 p* g- }
(1. S chool of Cl i nical Medici ne , L anz hou Uni versi t y , L anz hou , 730000 , Chi na;' {! j8 E! G7 o4 c& q+ v& I: o# z
2. Dep artment of I nf ect ion , T he Fi rst Hos pi tal of L anz hou Uni versi t y ,0 s/ m% a4 K8 `' P) g
L anz hou , 730000 , Chi na )
, G: G- Z% }! n) z( vAbstract : Ob j ectives To st udy the growt h pat tern and surface markers of i n vi t ro cult ured bone
6 y9 w) u7 ], i/ [; wmarrow mesenchymal stem cell s (MSCs) f rom liver cirrhosis patient s , provide new evidence for' Q( E5 L7 a2 v6 q% ]
t he choice of " seed cell s" for liver tissue engineer and lay basis for f urt her study of t he i n vi vo
, B% S# m& |7 f$ V- f7 u2 rregulatory mechanisms of t ransplanted autologous bone marrow MSCs in t he t reatment of liver3 a6 q8 I' m& k2 S! N w' a
failure. Met hods Bone marrow cell s were collected f rom 4 volunteer s aged f rom 30 to 40 wit h liver
! j) `- c$ w" l9 Zcirrhosis , MSCs were separated by a density gradient cent rif ugation , biological features of cul2
; p* K2 ~' k5 i, P! xt ured MSCs were observed under inverted microscope and their growt h curves were investigated.
) w( P8 I3 k% UFlow cytomet ry was used to detect surface antigenic markers of CD44 and CD45. Resul ts The la2, N y8 n1 L( ` a2 T1 `# z, x5 ~* T
tent p hase of the p rimary cultured cell s was 0 to 9 d followed by 10 to 19 d of logarithmical prolif2
3 Q9 M0 D$ T {- g' ]) aeration , and 20 to 24 d of platform growth. Passaged cells showed faster growt h t han primary
2 p+ j$ l' _. ^, g) i. ccell s , and t heir latent p hase was 4 to 24 h , logarit hmical proliferation p hase was 3 to 12 d and
: H% K! H/ e8 C# `# Yplatform growth was 13 to 15 d , MSCs expressed higher CD44 but lower CD45. ConclusionMSCs f rom liver cirrhosis patient s grow bet ter and have steady biological feat ures i n vi t ro , and
3 k R, K" o& {5 f/ t" o+ j/ Pmay be used as " seed cell s" for autologous bone marrow MSCs t MSCs f rom liver cirrhosis patient s grow bet ter and have steady biological feat ures i n vi t ro , and% J0 E& F% S) y0 d! d( k
may be used as " seed cell s" for autologous bone marrow MSCs t ransplantation and artificial liver . r/ {: i8 f8 F! `7 P" M7 [
Key words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy2% V1 |9 d/ u) D' S9 m, z
mal stem cell s ; t ransplantationransplantation and artificial liver .
# s4 m; y. `; M) _: ?6 k! F) Y1 [. i( u) v8 {( m5 K
附件是pdf全文。感谢大家支持，回复本贴就可以下载了# P: h( @3 B9 B% k% t
, {5 i; n" P+ V% k# |; F1 J/ T+ @/ D7 @( u; F( \( B( H
Key words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy2
$ R1 X/ a" i$ d; K/ t! Ymal stem cell s ; t ransplantation- h' s4 K1 e3 V: D! O) T" V
7 k! [ G) }" k/ o' s9 t+ M
附件是pdf全文。感谢大家支持，回复本贴就可以下载了
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发表于 2009-3-2 09:19
发表于 2009-6-5 21:03
