肝細胞の簡単な分離法の開発とその培養への応用

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  • A simple enzymatic method for isolation of rat hepatocytes and its application
  • カン サイボウ ノ カンタン ナ ブンリホウ ノ カイハツ ト ソノ バイヨウ
  • Effect of prednisolone on carbohydrate and protein metabolism in rat hepatocyte
  • 副腎皮質ホルモン投与時の培養肝細胞の代謝

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1) Procedure for isolation of rat hepatocytes: About 30ml of 0.05% bacterial collagenase in Ca-free Hanks' solution was directly injected into the aseptically removed liver tissue through tuberculine needle at a speed of 20ml/min. After a few minutes of injection, liver tissue was torn into small pieces by two forcepts and then filtered through a platinum sieve with 150 meshes. Filtrate was centrifuged several times at a very low speed (200rpm). The final residue was suspended in Eagle's minimum essential medium containing 20% fetal calf serum and used for primary culture. The yield of isolated hepatocyte was (7.86±4.24)×106/g wet liver and contamination by Kupffer cells was less than 1%. More than 95% of cells excluded trypan blue.<BR>2) Effect of prednisolone on gluconeogenesis and protein biosynthesis: Gluconeogenesis in cultured hepatocytes from alanine or lactate added to the culture media was enhanced by 100μg/dish of prednisolone. Contrary to it, incorporation of 14C-leucine or 3H-proline into the newly synthesized protein was inhibited by the same amount of prednisolone. Radioactivity of 3Hhydroxyproline was detected and the nature of the protein containing hydroxyproline was discussed.

収録刊行物

  • 肝臓

    肝臓 19 (4), 327-331, 1978

    一般社団法人　日本肝臓学会

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