本帖最后由 细胞海洋 于 2010-5-17 23:06 编辑 , ]* g; u" v4 ~7 U& n : x2 n9 c" \, I2 t n9 c安方梅1 , 邓永东2 , 赵正斌2 ,陈 红2 + R1 {% K5 w8 z. f/ M# [! Q(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) . \! t- ?& u& y J6 O$ E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 1 j+ U) {! y( ~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 7 `8 ~; |& @9 z, V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5 u' [1 r3 @5 i
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 7 ^- m" Q: y( V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! D3 |4 b) V! b7 D' D
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 ,+ I! N2 H1 p! C7 H8 r# w
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 . |9 }6 i; |; T* w2 {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。" T6 O& S7 \3 e# M* l" @: O: C6 S
关键词:肝硬化;骨髓间充质干细胞; 自体骨髓干细胞;移植3 {# [ H4 Z( ]0 V) @1 l
中图分类号:R33 文献标识码:A 3 a( V4 k9 Q; n# Y4 X4 OI n vit ro culture and growth characteristics of bone marrow , X% i' }# }6 T8 [; V5 n2 ?- tmesenchymal stemcells from liver cirrhosis patients : \5 J6 ]) E+ K/ ~& @+ jAN Fang2mei1 ,DEN G Yong2dong2 , ZHAO Zheng2bin2 , CHEN Hong2( u3 ^. D/ K; T4 q- ~
(1. S chool of Cl i nical Medici ne , L anz hou Uni versi t y , L anz hou , 730000 , Chi na; 6 i2 m% X7 Z @7 q: X: l2. Dep artment of I nf ect ion , T he Fi rst Hos pi tal of L anz hou Uni versi t y ,1 q& c" I7 X# Y# C4 l2 i; L2 J
L anz hou , 730000 , Chi na ) : s4 H% j3 x% y, m3 ~Abstract : Ob j ectives To st udy the growt h pat tern and surface markers of i n vi t ro cult ured bone( A! M; o" ~1 I" V/ I) O
marrow mesenchymal stem cell s (MSCs) f rom liver cirrhosis patient s , provide new evidence for & |8 A) k9 Q ]( ct 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 % z5 i$ V p5 \) t( I, lregulatory mechanisms of t ransplanted autologous bone marrow MSCs in t he t reatment of liver ( c# b; U- \; P8 P4 l0 y4 ^failure. Met hods Bone marrow cell s were collected f rom 4 volunteer s aged f rom 30 to 40 wit h liver; U* J+ L# b" I# L8 {: H
cirrhosis , MSCs were separated by a density gradient cent rif ugation , biological features of cul2- B4 _! k2 x* g
t ured MSCs were observed under inverted microscope and their growt h curves were investigated. 7 Z; N$ t9 s! }0 TFlow cytomet ry was used to detect surface antigenic markers of CD44 and CD45. Resul ts The la24 L2 X5 q* D( t' f v8 L
tent p hase of the p rimary cultured cell s was 0 to 9 d followed by 10 to 19 d of logarithmical prolif2# i" ^& F$ B& P- t& ^
eration , and 20 to 24 d of platform growth. Passaged cells showed faster growt h t han primary4 y/ n0 g& O- c4 u3 y7 O
cell s , and t heir latent p hase was 4 to 24 h , logarit hmical proliferation p hase was 3 to 12 d and \* B& F# Z$ P! M( Qplatform 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$ T x7 z/ ]( F8 z: m4 j# s3 I- T
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 , c, O! W% `0 p0 u% r2 emay be used as " seed cell s" for autologous bone marrow MSCs t ransplantation and artificial liver .. _. m& v4 h& N0 j f$ [; M* D5 V
Key words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy2 8 q: X" A8 O8 T8 \3 Fmal stem cell s ; t ransplantationransplantation and artificial liver .. C+ S3 C4 H" Z W: Q7 V% Q. z1 V
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Key words : cirrho sis ; bone marrow mesenchymal stem cell s ; autologous bone marrow mesenchy25 |% ~) x, C8 l x' ~
mal stem cell s ; t ransplantation$ G" M9 i# D: K1 \9 O+ n) Q R
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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 / s. J; r+ B$ F4 J0 c2 z" k4 |1 {
[attach]185[/attach]作者: ALLUL 时间: 2009-3-7 00:13