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The Isolation and Primary Culture of Calf Hepatocytes
Cai Zhang, Li-Ming Wang1, Guo-Wen Liu, Wei Su, Jian-Long Huang, Guang-Hong Xie, Zhe Wang*
Lab of Animal Nutritional and Metabolic Diseases & Toxicopathy, College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China; 1College of Animal Science & Technology, Sichuan Agricultural University, Yaan 625014,
Abstract: Calf hepatocytes were isolated by modified two steps collagenase perfusion and one step perfusion combined with tissue blocks collagenase digestion respectively， and cultured in hepatoZYME-SFM. The viability of cultured hepatocytes was assessed by trypan blue exclusion. The morphologic change of cultured hepatocytes was observed， and the concentrations of albumin， urea and lactate dehydrogenase (LDH) in the supernatant collected from different cultural period of better isolated system were examined. The results were as follows: hepatocytes obtained by modified two steps collagenase perfusion were intact and had a prosperous viability and an active function. The fluctuated changes of LDH leakage， albumin synthesis and urea level were displayed in one week， and the lesser LDH leakage and higher concentrations of albumin and urea were observed on the third day. It is concluded that the collagenase digestion method is feasible for isolation of hepatocytes. Calf hepatocytes have an optimal function on the third days of the primary culture.
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