Isolation and primary culture of bovine hepatocytes: albumin synthesis and adrenergic activation of glycogenolysis.

N. Nakagawa-Tosa, M. Morimatsu, Katsumi Mominoki, H. Nakatsuji, B. Syuto, M. Saito

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Abstract

We describe a technique for isolation and primary culture of bovine hepatocytes, and their metabolic characterization. Hepatocytes were isolated from the caudate lobe of bovine liver by perfusion with 0.25 mM ethylene-glycol tetraacetic acid and 0.05% collagenase. The viability and yield of the cells were 70-92% and 0.1-3.6 x 10(7) cells/g liver, respectively. When the isolated hepatocytes were cultured in Williams' medium E, they began to spread in 3 hr and formed monolayers in 24 hr. These monolayers were retained for at least 6 days. To monitor the metabolic activities specific to liver, synthesis and secretion of albumin were measured by labeling with [35S]-methionine and immunoprecipitation. This activity was low in isolated hepatocytes, but increased after culturing 1-3 days, and decreased again after 6 days. Glycogenolytic activity was also assessed by measuring glucose release to the medium by stimulation with epinephrine. The glycogenolytic response to epinephrine was also enhanced by culturing the hepatocytes 1-3 days, but was decreased after 6 days. Since the isolated bovine hepatocytes retained the liver-specific activities of albumin synthesis and glycogenolysis for several days in culture, these cells are useful for cellular and molecular studies on the functions of bovine liver.

Original languageEnglish
Pages (from-to)125-129
Number of pages5
JournalJournal of Veterinary Medical Science
Volume56
Issue number1
Publication statusPublished - Feb 1994
Externally publishedYes

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ASJC Scopus subject areas

  • Marketing
  • veterinary(all)

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