| 大鼠肝陷窝细胞分离、鉴定及纯度、活性分析 |
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| 引用本文： | 和春肖,谢来峰,王磊,樊晋宇,徐存拴. 大鼠肝陷窝细胞分离、鉴定及纯度、活性分析[J]. 河南科学, 2009, 27(9): 1072-1076 |
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| 作者姓名： | 和春肖 谢来峰 王磊 樊晋宇 徐存拴 |
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| 作者单位： | 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;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梯度离心结合免疫磁珠方法分离陷窝细胞（pit cell），用分化抗原簇8（cluster of differentiation 8，CD8）和分化抗原簇56（cluster of differentiation 56，CD56）的免疫组织化学方法定性、定位再生肝（regenerating liver，RD、分散的肝脏细胞及分离的肝陷窝细胞，用RT—PCR定量肝陷窝细胞的CD8和CD56mRNA，用蛋白免疫印迹定量肝陷窝细胞的CD8和CD56蛋白．初步证实，分离的肝陷窝细胞中，CD8和CD56阳性细胞占95％以上；PH后各时间点分离的陷窝细胞的CD8和CD56的mRNA量稳定，相应的蛋白量亦稳定．表明改进的分离陷窝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 陷窝细胞 免疫磁珠 标记蛋白 |
Isolation,Purification and Identification of Pit Cells in Rat Liver |
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| He Chunxiao,Xie Laifeng,Wang Lei,Fan Jinyu,Xu Cunshuan. Isolation,Purification and Identification of Pit Cells in Rat Liver[J]. Henan Science, 2009, 27(9): 1072-1076 |
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| Authors: | He Chunxiao Xie Laifeng Wang Lei Fan Jinyu Xu Cunshuan |
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| Affiliation: | 1. College of Life Science;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 done following Higgins et al,hepatic cells were scattered by two-step perfusion,and pit cells were isolated and purified by density gradient centrifugation with 60 % percoll and immunomagnetic beads. Immunocytochemistry method was used to qualitify and localize cluster of differentiation 8 (CD8)and cluster of differentiation 56(CD56)in liver tissue,the isolated hepatic cells,and the purified pit cells. The exrpressions of CD8 and CD56 were quantified using RT-PCR. The result... |
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| Keywords: | rat cell isolation and identification pit cells immunomagnetic beads marker proteins |
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