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本帖最后由 细胞海洋 于 2010-5-17 23:06 编辑
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T, j* K! i: G( O3 I+ @安方梅1 , 邓永东2 , 赵正斌2 ,陈 红2
1 H9 e$ e q7 c5 z& {(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)7 O4 i, @% I9 k- ~
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
' @, {! E8 b# l& V6 q0 C7 ^" K( s7 B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
2 U T/ _( z% o& `% R4 s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$ D1 n7 ?- |: M) m7 f( ]
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# h" e5 B& L& k8 v4 S4 Z5 _ M
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) j6 g z2 w3 @. W! r9 X1 e) y
d ,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 ,, w/ K, K7 Z! [* ^
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: c. B/ |9 l" B6 W: P. |: [# i, y1 |
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。8 D3 ^2 S O/ s# u" [
关键词:肝硬化;骨髓间充质干细胞; 自体骨髓干细胞;移植
1 @; s: K# x+ |1 ]& D4 g中图分类号:R33 文献标识码:A" C4 b( r0 G! x
I n vit ro culture and growth characteristics of bone marrow/ @3 l6 `0 p ?+ k" c" M
mesenchymal stemcells from liver cirrhosis patients/ @& G; ^. F0 P+ B( w/ F% N5 ^( C
AN Fang2mei1 ,DEN G Yong2dong2 , ZHAO Zheng2bin2 , CHEN Hong21 d0 l6 M' n. K* G3 s
(1. S chool of Cl i nical Medici ne , L anz hou Uni versi t y , L anz hou , 730000 , Chi na;
7 h: d0 }1 ^" W2. Dep artment of I nf ect ion , T he Fi rst Hos pi tal of L anz hou Uni versi t y ,: m% N* W0 e8 R' E7 C
L anz hou , 730000 , Chi na )+ Y7 A6 u% \1 e
Abstract : Ob j ectives To st udy the growt h pat tern and surface markers of i n vi t ro cult ured bone; q1 p' E6 d5 @
marrow mesenchymal stem cell s (MSCs) f rom liver cirrhosis patient s , provide new evidence for
! k; c% G8 \' y& }% ^5 ^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" A2 Y( L0 R1 O
regulatory mechanisms of t ransplanted autologous bone marrow MSCs in t he t reatment of liver
5 L. J7 \0 e; |& X4 X( J. T5 ^9 {failure. Met hods Bone marrow cell s were collected f rom 4 volunteer s aged f rom 30 to 40 wit h liver3 z' S. E* X1 M3 k, o( G
cirrhosis , MSCs were separated by a density gradient cent rif ugation , biological features of cul22 @1 D6 q' D3 i$ L
t ured MSCs were observed under inverted microscope and their growt h curves were investigated.3 G* n7 S: o5 P/ L
Flow cytomet ry was used to detect surface antigenic markers of CD44 and CD45. Resul ts The la2
3 K$ d9 G& l) |* V4 w0 itent p hase of the p rimary cultured cell s was 0 to 9 d followed by 10 to 19 d of logarithmical prolif2
. B& L6 C; l( G- x: W, oeration , and 20 to 24 d of platform growth. Passaged cells showed faster growt h t han primary
* e0 k) `: o1 s3 \cell s , and t heir latent p hase was 4 to 24 h , logarit hmical proliferation p hase was 3 to 12 d and) p. \5 \# e6 n) C
platform 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 , and0 H9 w0 F o' e8 R
may 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& ?. u0 f4 F+ h
may be used as " seed cell s" for autologous bone marrow MSCs t ransplantation and artificial liver .
8 p( X( n' c$ p1 U. hKey words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy2* H \. H7 b! n) v
mal stem cell s ; t ransplantationransplantation and artificial liver .
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7 e9 U2 z1 m1 p. b5 K x% b4 f0 |$ Y0 W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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) ?( P4 n( `" d5 ~+ zKey words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy2
5 K2 d. ~4 ]" S( @* C, Ymal stem cell s ; t ransplantation
, h$ P5 D' b# Z% b' a) ^& k9 g l# C ]1 Q" n8 ^8 S
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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