| 大鼠肝窦内皮细胞的分离、鉴定及纯度、活性分析 |
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| 引用本文： | 王泽,王磊,谢来峰,樊晋宇,徐存拴.大鼠肝窦内皮细胞的分离、鉴定及纯度、活性分析[J].河南科学,2009,27(8):930-934. |
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| 作者姓名： | 王泽 王磊 谢来峰 樊晋宇 徐存拴 |
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| 作者单位： | 1. 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
2. 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： | 河南省重大公益性科研计划项目 |
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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梯度离心和免疫磁珠分离肝窦内皮细胞（hepatic sinusoidal endothelial cell，SEC），用分化抗原簇14（cluster of differentiation 14，CD14）和内皮素-1（endothelin-1，ET-1）的免疫组织化学定性、定位再生肝（regenerating liver，RL）、分散的肝脏细胞及分离的窦内皮细胞，用RT—PCR定量窦内皮细胞的CDl1和ET-1的mRNA，用蛋白免疫印迹方法定量窦内皮细胞的CD14和ET-1蛋白．初步证实，分离的窦内皮细胞中CD14和ET-1阳性细胞占95％以上，从PH后各时间点分离的窦内皮细胞的CD14和ET-1mRNA量稳定，相应的蛋白量亦稳定．表明改进的分离窦内皮细胞方法具有收率和纯度高、活性好等特点，值得采用．
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| 关 键 词： | 大鼠 细胞分离和鉴定 窦内皮细胞 免疫磁珠 标记蛋白 |
Isolation,Purification and Identification of Sinusoidal Endothelial Cells in Rat Liver |
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| Wang Ze,Wang Lei,Xie Laifeng,Fan Jinyu,Xu Cunshuan.Isolation,Purification and Identification of Sinusoidal Endothelial Cells in Rat Liver[J].Henan Science,2009,27(8):930-934. |
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| Authors: | Wang Ze Wang Lei Xie Laifeng Fan Jinyu Xu Cunshuan |
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| Institution: | 1. College of Life Sciences;Henan Normal University;Xinxiang 453007;Henan China;2. Key Laboratory for the Research of Cell Differentiation and Regulation;Henan China |
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| Abstract: | Rat 2/3 hepatectomy(PH)model was made following Higgins et al.,hepatic cells were scattered by two-step perfusion,and sinusoidal endothelial cells were isolated by density gradient centrifugation with 60 % percol1 and immuno-magnetic beads. Immunocytochemistry method was used to qualitify and localize cluster of differentiation 14(CD14) and endothelin-1(ET-1) in liver tissue,the isolated hepatic cells,and the purified sinusoidal endothelial cells. The expressions of CD14 and ET-1 were quantified using RT-PC... |
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| Keywords: | rat cell isolation and identification sinusoidal endothelial cells(SEC) immuno-magnetic bead marker protein |
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