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当前位置: 首页 > 文章 > 广西瑶山亚种树鼩原代肝细胞的分离、纯化、培养及鉴定 黑龙江畜牧兽医 2017 (7) 42-45+290
Position: Home > Articles > Isolation,purification,culture and identification of primary hepatocytes in Tupaia belangeri yaoshanensis from Guangxi Heilongjiang Animal Science and Veterinary Medicine 2017 (7) 42-45+290

广西瑶山亚种树鼩原代肝细胞的分离、纯化、培养及鉴定

作  者：
刘金花;农微;黄正团;运晨霞;夏钰淇;冷静
单  位：
广西中医药大学
关键词：
树鼩;肝细胞;分离;纯化;培养;PAS染色;免疫细胞化学
摘  要：
为了探讨瑶山亚种树原代肝细胞分离、培养及鉴定方法,试验对传统的两步原位胶原酶灌注法进行了优化,先将持续灌注法改为间断夹闭下腔静脉快速灌注法,再通过严格控制胶原酶的消化时间来分离提取树肝细胞,最后应用percoll分离液梯度低速离心纯化树肝细胞,采用PAS染色法和免疫细胞化学方法对树肝细胞进行综合鉴定。结果表明:分离得到的树肝细胞产量高、活力好;经PAS染色鉴定,发现树肝细胞内糖原被染成紫红色颗粒或片状;免疫细胞化学染色观察到树肝细胞胞浆内被染成棕黄色。说明成功分离培养树鼩肝细胞并对其进行鉴定,可用于进一步研究。
译  名：
Isolation,purification,culture and identification of primary hepatocytes in Tupaia belangeri yaoshanensis from Guangxi
作  者：
LIU Jinhua;NONG Wei;HUNG Zhengtuan;YUN Chenxia;XIA Yuqi;LENG Jing;Guangxi University of Chinese Medicine;
关键词：
Tupaia belangeri;;hepatocyte;;isolation;;purification;;culture;;PAS staining;;immunocytochemistry
摘  要：
To investigate the methods of isolation,cultivation and identification of primary hepatocytes in Tupaia belangeri yaoshanensis,the conventional two-step collagenase perfusion in situ method was optimized.A continuous perfusion was replaced by a rapid perfusion of intermittent occlusion of inferior vena cava,and then the collagenase digestion time was strictly controlled to isolate and extract the hepatocytes in Tupaia belangeri yaoshanensis.The hepatocytes were purified by percoll gradient centrifugation at low speed,and then were comprehensively identified using PAS staining and immunocytochemistry.The results showed that the hepatocytes were isolated and obtained,which had high yield and good viability; it was found that the glycogens in the hepatocytes were stained purple granules or flakes by PAS staining,and the cytoplasm in the hepatocytes was stained brown by immunocytochemistry staining.The results indicate that the primary hepatocytes in Tupaia belangeri yaoshanensis,which can be used for further research.

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