...
首页> 外文期刊>The Veterinary Journal >An updated method for the isolation and culture of primary calf hepatocytes
【24h】

An updated method for the isolation and culture of primary calf hepatocytes

机译:一种更新的分离和培养小牛肝细胞的方法

获取原文
获取原文并翻译 | 示例
   

获取外文期刊封面封底 >>

       

摘要

Primary hepatocytes are commonly used during in vitro studies, but care must be taken with isolation and culture of the cells to ensure their viability. In this study, hepatocytes were isolated from the liver (caudate process) of a newborn calf by the collagenase perfusion and digestion method. The trypan blue exclusion method was used to determine total cell number and the survival rate of hepatocytes, while hepatocyte function was assessed by measuring lactate dehydrogenase. albumin and urea in culture medium supernatants at 24, 48, 72, 96, 120, 144 and 168 h. Results showed that the number of viable cellsig of liver (wet weight) averaged 1.12 x 10(7) cells/g, with an average hepatocyte viability of 85.7% (range 83-92%). After 48 h of culture, the hepatocytes solidly adhered to the well culture plate and were spread in an epithelioid shape, with clear cell boundaries between the cells and biliary ductule-like structures formed which persisted for up to 10 days. Hepatocyte function was optimal at 72 h after isolation and culture. This simple and economical procedure for the isolation and culture of viable cells may be useful for in vitro bovine hepatocyte studies
机译:原代肝细胞通常用于体外研究,但必须小心分离和培养细胞以确保其生存能力。在这项研究中,通过胶原酶灌注和消化方法从新生小牛的肝脏(尾状过程)中分离出肝细胞。锥虫蓝排除法用于确定总细胞数和肝细胞存活率,而肝细胞功能通过测量乳酸脱氢酶来评估。在24、48、72、96、120、144和168小时的培养基上清液中加入白蛋白和尿素。结果显示,肝脏的活细胞数(湿重)平均为1.12 x 10(7)细胞/ g,平均肝细胞活力为85.7%(范围为83-92%)。培养48小时后,肝细胞牢固地粘附在孔培养板上,并呈上皮状扩散,在细胞之间形成清晰的细胞边界,并形成胆小管样结构,持续长达10天。分离和培养后72 h,肝细胞功能最佳。这种简单且经济的分离和培养活细胞的方法可能对体外牛肝细胞研究有用

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号