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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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安方梅1 , 邓永东2 , 赵正斌2 ,陈 红24 F. l; w" E% A* l+ }: Z% y
(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)
1 q$ J/ w' L0 _; X% @3 y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
$ P3 O1 N5 ^+ H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+ i& V$ v6 a4 y* J6 {# N
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
+ _* q6 x' p+ ~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
5 | g% l6 L3 l* e/ t! |# N) @" r; n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10～194 s( j, D1 I1 E# ?
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 ,
- U1 c1 w* {# D5 Z( ~2 L& U13～15 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- r2 k% J' C, G4 B. X/ j4 L- A
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 _! q6 o7 C* [9 [7 ?
关键词:肝硬化;骨髓间充质干细胞; 自体骨髓干细胞;移植
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I n vit ro culture and growth characteristics of bone marrow; w9 a& ~7 \) I
mesenchymal stemcells from liver cirrhosis patients
/ u- K. c0 Y$ \; qAN Fang2mei1 ,DEN G Yong2dong2 , ZHAO Zheng2bin2 , CHEN Hong2
) x# l0 ~- s) B3 Z(1. S chool of Cl i nical Medici ne , L anz hou Uni versi t y , L anz hou , 730000 , Chi na;* T$ m4 O; L8 d9 t
2. Dep artment of I nf ect ion , T he Fi rst Hos pi tal of L anz hou Uni versi t y ,, t3 C7 u8 F B J
L anz hou , 730000 , Chi na )2 V% G2 u! u* g" ]8 O% d
Abstract : Ob j ectives To st udy the growt h pat tern and surface markers of i n vi t ro cult ured bone
: K Y, Y8 Z _4 S& omarrow mesenchymal stem cell s (MSCs) f rom liver cirrhosis patient s , provide new evidence for' A' z2 y) Q# U: \2 M* ~. @
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
- X: `2 G T0 K$ Y9 i. z6 `regulatory mechanisms of t ransplanted autologous bone marrow MSCs in t he t reatment of liver" l' f6 L1 R1 r9 w- Q% _
failure. Met hods Bone marrow cell s were collected f rom 4 volunteer s aged f rom 30 to 40 wit h liver
3 ~' g% ~3 G, ncirrhosis , MSCs were separated by a density gradient cent rif ugation , biological features of cul2
$ Y- O" x9 C& Z- N1 ?+ V8 T6 yt ured MSCs were observed under inverted microscope and their growt h curves were investigated.
' }3 v/ e( e' X0 QFlow cytomet ry was used to detect surface antigenic markers of CD44 and CD45. Resul ts The la2, i' @/ C: f' j' o9 N1 C
tent p hase of the p rimary cultured cell s was 0 to 9 d followed by 10 to 19 d of logarithmical prolif2
. b# b- p8 ?! d h6 J( Weration , and 20 to 24 d of platform growth. Passaged cells showed faster growt h t han primary* {6 \# Y. i/ g: I2 l1 _
cell s , and t heir latent p hase was 4 to 24 h , logarit hmical proliferation p hase was 3 to 12 d and
) n% u- O8 `' D3 v. Iplatform 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
% ^+ z0 u! K* g6 g5 i) N& \( {. d3 X3 [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 , and3 K' P1 c4 \5 b
may be used as " seed cell s" for autologous bone marrow MSCs t ransplantation and artificial liver .
" g2 t0 G7 `) E0 N, j$ n! YKey words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy2, k) a$ ?+ \; o0 q
mal stem cell s ; t ransplantationransplantation and artificial liver .
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3 u, S; q, o. B9 B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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Key words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy2
{1 X/ _; q% h0 o* R2 E0 ~2 xmal stem cell s ; t ransplantation
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. r8 r" p: L" g2 F4 t+ K# K! ~附件是pdf全文。感谢大家支持，回复本贴就可以下载了; P7 v* g! _6 |3 u
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发表于 2009-3-2 09:19
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