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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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) i2 R7 V8 Q: P9 V7 I$ V j安方梅1 , 邓永东2 , 赵正斌2 ,陈 红2
0 D4 C6 x9 _+ Y) _- A(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)
( k7 `0 j7 i! T8 w# V. U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
! h5 h! w5 \% ^% O3 a5 C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
% J1 ~2 C& h9 K- K1 D* F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% ~$ s3 b; O) X3 n% J' A: \1 H
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, L4 y+ i# _- k$ z+ h
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
! e, t) v% W5 G. E' n! ?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 ,- Y/ x5 r, Y" K6 ~4 N
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0 e& A. `( z4 b+ S6 O2 C' W
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。
% K; k9 [/ x7 ?9 X关键词:肝硬化;骨髓间充质干细胞; 自体骨髓干细胞;移植
8 y9 z! L* l- L中图分类号:R33 文献标识码:A
$ ?# m# E* x, n. H CI n vit ro culture and growth characteristics of bone marrow4 @( X/ ^+ w' F+ u' Z3 m% A
mesenchymal stemcells from liver cirrhosis patients3 y, r1 h: B$ }4 x/ R/ c1 g
AN Fang2mei1 ,DEN G Yong2dong2 , ZHAO Zheng2bin2 , CHEN Hong28 }* P( B3 D2 s; D0 e
(1. S chool of Cl i nical Medici ne , L anz hou Uni versi t y , L anz hou , 730000 , Chi na;3 b2 E/ J. Q" _, x( v
2. Dep artment of I nf ect ion , T he Fi rst Hos pi tal of L anz hou Uni versi t y ,
$ O% |# D6 s h6 o5 UL anz hou , 730000 , Chi na )9 Z9 o3 G) m3 B: @) q
Abstract : Ob j ectives To st udy the growt h pat tern and surface markers of i n vi t ro cult ured bone! S, \$ w/ t% [7 n3 d5 j& ^
marrow mesenchymal stem cell s (MSCs) f rom liver cirrhosis patient s , provide new evidence for
9 n6 D; q8 s, K' ]. n& v! yt 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
+ o8 H+ X4 ]" R* A. [/ f$ z0 hregulatory mechanisms of t ransplanted autologous bone marrow MSCs in t he t reatment of liver) e/ W# h$ u9 Z( `: |1 P
failure. Met hods Bone marrow cell s were collected f rom 4 volunteer s aged f rom 30 to 40 wit h liver
% S5 \# a; M7 }cirrhosis , MSCs were separated by a density gradient cent rif ugation , biological features of cul2
2 ~( s9 r/ P9 I. H. {t ured MSCs were observed under inverted microscope and their growt h curves were investigated.
, r, q) O5 V: r* I6 J5 M d9 {Flow cytomet ry was used to detect surface antigenic markers of CD44 and CD45. Resul ts The la24 r) q3 F5 v7 z) U9 A: p1 X
tent p hase of the p rimary cultured cell s was 0 to 9 d followed by 10 to 19 d of logarithmical prolif2+ |- u% q& I* w) Y) J* F& a5 [
eration , and 20 to 24 d of platform growth. Passaged cells showed faster growt h t han primary
, D; ~& T" K2 B! zcell s , and t heir latent p hase was 4 to 24 h , logarit hmical proliferation p hase was 3 to 12 d and8 U$ [! |$ u0 K8 A
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 , and
8 ~/ z, n! x0 p! Dmay 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
2 h6 C. Y/ |8 u* {! ^% @. }4 Dmay be used as " seed cell s" for autologous bone marrow MSCs t ransplantation and artificial liver .
# l) L0 j+ S) a5 pKey words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy2
& O- b+ s* F! Mmal stem cell s ; t ransplantationransplantation and artificial liver .! x: n" O. g* _# ^3 x6 m8 ^
' P/ M1 @, V& K附件是pdf全文。感谢大家支持，回复本贴就可以下载了8 L9 Z4 M: f0 t
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# N* c5 _4 e/ N; W8 yKey words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy2! }- q; h/ P: m
mal stem cell s ; t ransplantation
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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
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