| 大鼠正常肝和再生肝的肝细胞分离、鉴定及纯度、活性分析 |
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| 引用本文： | 王文博,谢来峰,王望,王磊,徐存拴.大鼠正常肝和再生肝的肝细胞分离、鉴定及纯度、活性分析[J].河南科学,2008,26(12). |
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| 作者姓名： | 王文博 谢来峰 王望 王磊 徐存拴 |
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| 作者单位： | 1. 中国海洋大学,水产学院,青岛,266100;河南省-科技部共建细胞分化调控重点实验室,河南,新乡,453007
2. 河南师范大学,生命科学学院,河南,新乡,453007;河南省-科技部共建细胞分化调控重点实验室,河南,新乡,453007 |
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| 基金项目： | 国家973计划前期研究专项 |
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| 摘 要： | 按Higgins等方法制作大鼠2/3肝切除(parital hepatectomy,PH)模型,用两步灌流法分散肝脏细胞,用60%Percoll密度梯度离心分离肝细胞,用免疫组织化学方法定性、定位再生肝(regenerating liver,RL)、分散的肝脏细胞及分离的肝细胞中白蛋白(albumin,ALB)和葡萄糖-6-磷酸酶(glucose-6-phosphatase,G-6-P),用蛋白免疫印迹方法定量肝细胞的ALB和G-6-P,用RT-PCR定量肝细胞的ALB和G-6-P mRNA.初步证实纯化的细胞中,ALB和G-6-P阳性细胞占96%以上;PH后各时间点纯化的肝细胞的ALB和G-6-P mRNA量稳定,相应的蛋白量亦稳定.表明改进后的分离肝细胞方法具有收率高、纯度高、活性好等特点.
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| 关 键 词： | 大鼠 细胞分离和鉴定 肝细胞 基因表达 标记蛋白 |
Isolation,Purity and Identification of Hepatocytes in Rat Normal Liver and Regenerating Liver |
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| Wang Wenbo,Xie Laifeng,Wang Wang,Wang Lei,Xu Cunshuan.Isolation,Purity and Identification of Hepatocytes in Rat Normal Liver and Regenerating Liver[J].Henan Science,2008,26(12). |
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| Authors: | Wang Wenbo Xie Laifeng Wang Wang Wang Lei Xu Cunshuan |
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| Institution: | Wang Wenbo1,3,Xie Laifeng2,Wang Wang2,Wang Lei2,Xu Cunshuan2,3(1. College of Fisheries,Ocean University of China,Qingdao 266100,China,2. College of Life Sciences,Henan Normal University,Xinxiang 453007,Henan China,3. HNP/MOST Co-construction Key Laboratory for Cell Differentiation Regulation,Henan China) |
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| Abstract: | Rat 2/3 hepatectomy model was made by the method of Higgins et al.,hepatic cells were scattered by two-step perfusion,and hepatocytes were isolated and pruified by density gradient centrifugation with 60 % percoll. Immunocytochemistry method was used to qualitify and localize albumin(ALB) and glucose-6-phosphatase(G-6-P) in liver tissue,the dispersed hepatic cells,and the purified hepatoctyes. The exrpressions of alb and G-6-P were quantified using RT-PCR. The results primarily showed that ALB/G-6-P postive... |
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| Keywords: | rat cell isolation and identification hepatocyte gene expression marker protein |
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