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本帖最后由 细胞海洋 于 2010-5-17 23:06 编辑 " E$ R+ g3 U7 q' z% o1 q
% u7 ~) F0 i; Q% s6 C, ]安方梅1 , 邓永东2 , 赵正斌2 ,陈 红2
5 l8 i4 v! p; D3 x. 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)
) R6 Z, e* I* M. _# a5 C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
$ S/ U2 |" H$ ~" j4 |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
) o$ a B7 ]$ W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7 ^6 `& G& W3 A& y' j/ ^8 Z
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+ B/ Y# l7 t# S) ^
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10～193 `( L# b; Q& A1 O- T' U! r7 p7 n& S
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 ,
: _9 f2 m& M7 o( B+ u% u6 M4 Z13～15 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
* a0 v4 u( G0 x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。
! u" u6 I" Z- c/ k/ u关键词:肝硬化;骨髓间充质干细胞; 自体骨髓干细胞;移植
; G9 w" z3 w! A5 t中图分类号:R33 文献标识码:A
& u R( i' w5 t! l) f5 p+ e) hI n vit ro culture and growth characteristics of bone marrow
8 E# b) `; R9 ?mesenchymal stemcells from liver cirrhosis patients
( F& M$ Z: n! [; \: |AN Fang2mei1 ,DEN G Yong2dong2 , ZHAO Zheng2bin2 , CHEN Hong2
' x' E* e# D& u3 `$ v! O+ }(1. S chool of Cl i nical Medici ne , L anz hou Uni versi t y , L anz hou , 730000 , Chi na;3 z( g5 B7 e3 w" e* e0 D0 O4 S1 E# h( F. z
2. Dep artment of I nf ect ion , T he Fi rst Hos pi tal of L anz hou Uni versi t y ,* U3 n" p4 Q% p+ H: ?
L anz hou , 730000 , Chi na )! J$ H: o: S2 l" s- ?% W- |$ x
Abstract : Ob j ectives To st udy the growt h pat tern and surface markers of i n vi t ro cult ured bone
. u6 f5 h8 k0 R3 I3 kmarrow mesenchymal stem cell s (MSCs) f rom liver cirrhosis patient s , provide new evidence for
) [4 Y( F8 f; J2 Xt 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: P/ U% x8 s! K* Q$ b: Q- S
regulatory mechanisms of t ransplanted autologous bone marrow MSCs in t he t reatment of liver0 _' _9 v% W& _. H! K. R1 n
failure. Met hods Bone marrow cell s were collected f rom 4 volunteer s aged f rom 30 to 40 wit h liver
$ l! g4 s) `. V' ~! u" Zcirrhosis , MSCs were separated by a density gradient cent rif ugation , biological features of cul2
; ~( K2 G! m+ g- D0 X0 M- Wt ured MSCs were observed under inverted microscope and their growt h curves were investigated.
- H& _) q; I0 S3 \# Y% X" qFlow cytomet ry was used to detect surface antigenic markers of CD44 and CD45. Resul ts The la2
& X- L" @5 P: P* _tent p hase of the p rimary cultured cell s was 0 to 9 d followed by 10 to 19 d of logarithmical prolif26 J' [& v" ]: H0 U
eration , and 20 to 24 d of platform growth. Passaged cells showed faster growt h t han primary
& s# q) E/ ~- l2 o! ]( ~ k" Xcell s , and t heir latent p hase was 4 to 24 h , logarit hmical proliferation p hase was 3 to 12 d and
- @5 Y% @+ P7 g' Eplatform 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# J. h1 W5 C! j, w2 R# ~& U
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
# Y$ \/ r" E# [# _7 _3 Mmay be used as " seed cell s" for autologous bone marrow MSCs t ransplantation and artificial liver .
' t, T% q3 d3 P$ KKey words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy24 F1 d6 ~1 a! Q! h3 p
mal stem cell s ; t ransplantationransplantation and artificial liver .
3 [( W, V$ L' K: J( ^1 f
- }+ d" p. ?0 }' N! }) W" Q7 ]附件是pdf全文。感谢大家支持，回复本贴就可以下载了3 Q0 q; o% O, r! }- i
! X: U- X( f. |4 q
9 d1 c3 k6 E* J' O6 Z& A3 iKey words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy2/ D" }5 t: G8 e
mal stem cell s ; t ransplantation
: H/ |! P# j9 r$ O: |9 z$ v$ `; I C$ O1 ^- U7 \
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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