کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2464343 1111791 2012 4 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
An updated method for the isolation and culture of primary calf hepatocytes
موضوعات مرتبط
علوم زیستی و بیوفناوری علوم کشاورزی و بیولوژیک علوم دامی و جانورشناسی
پیش نمایش صفحه اول مقاله
An updated method for the isolation and culture of primary calf hepatocytes
چکیده انگلیسی

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 cells/g of liver (wet weight) averaged 1.12 × 107 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.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: The Veterinary Journal - Volume 191, Issue 3, March 2012, Pages 323–326
نویسندگان
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