肝硬化患者骨髓间充质干细胞的体外培养及生长特性

cdk1

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安方梅1 , 邓永东2 , 赵正斌2 ,陈　红2/ l( |& h( O: B8 C% v$ G# R( s. x3 ~# E" E
(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); \9 {1 L9 a9 m, M9 g, p
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
/ a" p' L  Q; G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# x# {) u1 ?0 [! x+ 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, ]2 ]. C9 b1 S# n& d2 ?! c; ]
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 H% |; B! s! @& M1 i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) i( E7 ~# F1 n6 Td ,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 ,
7 W- N$ `  S$ g13～15 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5 Z; Y3 w& u6 k8 i/ u  f9 @0 {& R6 V4 t
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。
! l, L1 w/ j4 T' w2 J3 c关键词:肝硬化;骨髓间充质干细胞; 自体骨髓干细胞;移植/ P* q* r" j0 {; I( K
中图分类号:R33 　　　　　　　　文献标识码:A! k  p' X5 K0 {" N# ?+ B' M1 q& S$ X
I n vit ro culture and growth characteristics of bone marrow
) r& g: z. F/ [3 Wmesenchymal stemcells from liver cirrhosis patients- N  H: Q: q  O* [* @
AN Fang2mei1 ,DEN G Yong2dong2 , ZHAO Zheng2bin2 , CHEN Hong2
9 C/ O& H' {! k(1. S chool of Cl i nical Medici ne , L anz hou Uni versi t y , L anz hou , 730000 , Chi na;
2 N, H1 S; v( M+ X$ i/ |4 i2. Dep artment of I nf ect ion , T he Fi rst Hos pi tal of L anz hou Uni versi t y ,: V% `0 E, O2 H- G" k! |6 R
L anz hou , 730000 , Chi na )
2 B5 n% t* b- W( k. kAbstract : Ob j ectives To st udy the growt h pat tern and surface markers of i n vi t ro cult ured bone
9 D$ Q2 W; B5 imarrow mesenchymal stem cell s (MSCs) f rom liver cirrhosis patient s , provide new evidence for2 c. J; Y1 E2 D8 |/ Z0 y3 A. k
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! P0 F! [- c9 v7 O8 [7 M
regulatory mechanisms of t ransplanted autologous bone marrow MSCs in t he t reatment of liver7 _2 P0 t; r9 Y2 M& S5 j( n
failure. Met hods Bone marrow cell s were collected f rom 4 volunteer s aged f rom 30 to 40 wit h liver& F( |' D6 [: z1 Q: \0 d6 H8 H
cirrhosis , MSCs were separated by a density gradient cent rif ugation , biological features of cul2' y3 z( L$ W9 f7 s7 }$ \6 y
t ured MSCs were observed under inverted microscope and their growt h curves were investigated.
2 |. G! Q6 W. ~2 c" w* F( iFlow cytomet ry was used to detect surface antigenic markers of CD44 and CD45. Resul ts The la2
+ t  B" d1 a, m# Ftent p hase of the p rimary cultured cell s was 0 to 9 d followed by 10 to 19 d of logarithmical prolif2
+ a. J3 I9 C& v; _% Jeration , and 20 to 24 d of platform growth. Passaged cells showed faster growt h t han primary
7 j2 ?( T7 i; p" gcell s , and t heir latent p hase was 4 to 24 h , logarit hmical proliferation p hase was 3 to 12 d and# H' ~& G/ |- X3 ?  a7 u
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
: n& n0 O4 m+ F6 @" }$ {7 W) kmay 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 J/ I5 z% @6 c0 ], `may be used as " seed cell s" for autologous bone marrow MSCs t ransplantation and artificial liver .  {% h& B5 i/ _* |+ U, p
Key words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy21 ]+ s7 p! Q- [+ I
mal stem cell s ; t ransplantationransplantation and artificial liver .
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- R5 r( J8 k! hKey words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy2
/ E% p4 C1 n# j% \, i" Y# _' Bmal stem cell s ; t ransplantation1 j0 x7 Q2 C2 Z7 C4 `8 g

& M5 n9 w1 n# [5 @, J) F+ T附件是pdf全文。感谢大家支持，回复本贴就可以下载了5 |8 G9 J) x' `6 [: ~% P: P) o

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ALLUL

非常感谢大家

horst

真是好资料，很有帮助，谢谢了

sims

谢谢楼主！！！

fountaincs

谢谢楼主

ligangcangxb

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barton_nyd

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hualin840518

xiexie

medusa75

十分感谢