| Optiprep法分离大鼠肝星状细胞的实验研究 |
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| 引用本文： | 余盼攀,张胜初,季世强,苏龙丰,张启瑜.Optiprep法分离大鼠肝星状细胞的实验研究[J].温州医学院学报,2011,41(6):511-514. |
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| 作者姓名： | 余盼攀 张胜初 季世强 苏龙丰 张启瑜 |
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| 作者单位： | 温州医学院附属第一医院,肝胆外科,浙江,温州,325035 |
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| 基金项目： | 浙江省外科学重中之重学科基金资助项目 |
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| 摘 要： | 目的：建立一种合理、经济、便捷的大鼠原代肝星状细胞（hepatic stellate cells,HSC）分离方法。方法：运用改良肝脏原位灌注D-Hanks后,改用Ⅳ型胶原酶和DNA酶Ⅰ离体灌注,10%、16%Optiprep密度梯度离心分离大鼠肝星状细胞,台盼蓝排斥试验鉴定细胞活力,α-SMA免疫细胞化学法鉴定细胞纯度。结果：我们采用Optiprep密度梯度离心法成功分离出大鼠HSC,且活力、纯度均达到研究要求,HSC得率约为2×107/肝,细胞存活率为95%以上,纯度为90%以上。结论：本方法分离大鼠HSC合理、经济、便捷。
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| 关 键 词： | 肝星状细胞 Optiprep 分离 大鼠 |
Experimental study on isolation of hepatic stellate cell in rats by Optiprep |
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| YU Panpan,ZHANG Shengchu,JI Shiqiang,SU Longfeng,ZHANG Qiyu.Experimental study on isolation of hepatic stellate cell in rats by Optiprep[J].Journal of Wenzhou Medical College,2011,41(6):511-514. |
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| Authors: | YU Panpan ZHANG Shengchu JI Shiqiang SU Longfeng ZHANG Qiyu |
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| Affiliation: | YU Panpan,ZHANG Shengchu,JI Shiqiang,SU Longfeng,ZHANG Qiyu.Department of Hepatobiliary Surgery,the First Hospital of Wenzhou Medical College,Wenzhou,325035 |
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| Abstract: | Objective: To established a reasonable,ecnomic and convenient method of isolating hepatic stellate cells(HSC) in rats.Methods: The method for isolation of rat hepatic stellate cells was established with D-Hanks in situ liver recirculating perfusion and then isolated perfusion with Type IV collagenase and DNaseⅠand continuous density gradient centrifugation by 10%,16% Optiprep Solution.The viability of the isolated cells was deter-mined with trypan blue staining assay.The HSC were identified by immunocytochemical staining of α-SMA.Results: the rat HSC were successfully isolated and the viability and purity were reached the requirement.The average number of HSC from a single rat liver was 2×107,with viability over 95%,purity over 90%.Conclusion: This method for isolating HSC is reasonable,ecnomic and convenient. |
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| Keywords: | hepatic stellate cells Optiprep isolation rats |
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