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| 成人原代肝细胞的分离、培养及冻存 | |
| 引用本文： | 杭化莲,张磊,施晓雷,卞建民,丁义涛. 成人原代肝细胞的分离、培养及冻存[J]. 世界华人消化杂志, 2011, 0(19) |
| 作者姓名： | 杭化莲 张磊 施晓雷 卞建民 丁义涛 |
| 作者单位： | 南京医科大学附属南京第一医院普外科;南京大学医学院附属鼓楼医院肝胆外科; |
| 摘 要： | 目的:建立一种稳定的成人原代肝细胞分离、培养、冻存方法,为肝细胞移植、生物人工肝支持系统治疗急慢性肝病以及肝细胞体外应用模型提供潜在的肝细胞资源.方法:20例供肝采用离体两步胶原酶灌注技术分离成人原代肝细胞.选择7个不同的预培养时间点(2、6、12、24、36、48和72h),分离所获肝细胞按上述不同预培养时间在4℃人无血清培养基(HepatoZYME-SFM)中培养,然后收集预培养肝细胞转移到含100mL/L胎牛血清和DMSO的HepatoZYME-SFM中,再立即放入-80℃异丙醇冷冻盒过夜,次日投入液氮.分析比较各肝细胞在解冻后细胞活力率、贴壁率、白蛋白分泌及尿素合成.结果:在部分肝叶切除后使用离体两步胶原酶灌流技术分离所得肝细胞活率和贴壁率分别是75.0%±4.6%和72.0%±6.0%.4℃预培养12或24h被证明是最适预培养时间,这两个时间点的白蛋白分泌高于其他时间点(P<0.05).与立即冷冻组相比较,预培养12或24h肝细胞解冻后活力(61.4%±4.8%,62.0%±5.6%vs53.4%±4.2%)、贴壁率(63.2%±5.8%,62.6%±3.6%vs55.2%±4.6%)、白蛋白分泌及... |
| 关 键 词： | 人原代肝细胞 肝细胞分离 冷冻保存 |
Isolation,culture and cryopreservation of adult human hepatocytes |
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| Hua-Lian Hang,Lei Zhang,Jian-Min Bian. Isolation,culture and cryopreservation of adult human hepatocytes[J]. World Chinese Journal of Digestology, 2011, 0(19) | |
| Authors: | Hua-Lian Hang Lei Zhang Jian-Min Bian |
| Affiliation: | Hua-Lian Hang,Lei Zhang,Jian-Min Bian,Department of General Surgery,Nanjing First Hospital,Nanjing Medical University,Nanjing 210006 Jiangsu Province,China Xiao-Lei Shi,Yi-Tao Ding,Department of Hepatobiliary Surgery,Affiliated Drum Tower Hospital of Nanjing University Medical School,Nanjing 210008,Jiangsu Province,China |
| Abstract: | AIM:To establish a stable method for the iso- lation,culture and cryopreservation of adult primary hepatocytes to provide a potential hepatocyte resource for the treatment of acute and chronic liver diseases using hepatocyte transplantation and bioartificial liver support systems,and for the use of hepatocytes as an in vitro model of the liver. METHODS:Adult hepatocytes were isolated from 20 separate donors using a two-step extra-corporeal collagenase perfusion technique.The hepatocytes were preincubated in... |
| Keywords: | Human hepatocytes Hepatocyte isolation Cryopreservation |
| 本文献已被 CNKI 等数据库收录！ | |