ヒト肝細胞の3次元培養スフェロイドモデルの新展開  [in Japanese] Further investigation of 3D culture spheroid models of human hepatocytes  [in Japanese]

Access this Article

Search this Article

Author(s)

    • 荻原 琢男 Ogihara Takuo
    • 高崎健康福祉大学 薬学部 薬学科 生物薬剤学研究室 Laboratory of Biopharmaceutics, Department of Pharmacology, Faculty of Pharmacy, Takasaki University of Health and Welfare
    • 細野 麻友 Hosono Mayu
    • 高崎健康福祉大学 薬学部 薬学科 生物薬剤学研究室 Laboratory of Biopharmaceutics, Department of Pharmacology, Faculty of Pharmacy, Takasaki University of Health and Welfare
    • 小島 肇 Kojima Hajime
    • 国立医薬品食品衛生研究所 安全性生物試験研究センター 安全性予測評価部 Division of Risk Assessment, Biological Safety Research Center, National Institute of Health Sciences

Abstract

<p>3次元(3D)培養肝細胞スフェロイドが,薬物代謝,毒性および酵素誘導の新しい評価として注目されている.ヒト肝細胞をスフェロイド用プレートに播種し細胞形態およびアルブミン分泌を検討したところ,肝細胞スフェロイドは播種後21日間維持されることが確認された.このスフェロイドに薬物を曝露した結果,第Ⅰ相および第Ⅱ相酵素による連続的な代謝反応およびヒト特有の代謝反応が認められた.また,このスフェロイドを用いて肝毒性を評価した結果,アルブミン分泌の50%阻害濃度は従来評価法よりも低く,従来評価法が肝毒性を過小評価している可能性が示唆された.さらに,このスフェロイドを用いて各種シトクロムP450(CYP)の誘導能を調べたところ,各種CYPのmRNA発現量および活性は有意に増加した.以上より,この3D肝細胞スフェロイドは,薬物代謝過程の追跡,肝毒性および酵素活性誘導の長期試験に適していることが示された.</p>

<p>Three-dimensional (3D) cultured hepatocyte capable of maintaining liver-specific function in an in vivo state over a relatively long period of time have drawn attention as a new method for evaluating the metabolic process, hepatotoxicity and enzyme induction potential of drugs. When human hepatocytes were seeded on a plate for spheroid formation, and cell morphology and albumin secretion were examined, hepatocyte spheroid was stably maintained for at least 21 days after seeding. As a result of drug exposure to this spheroid, sequential metabolic reactions by Phase I and Phase II enzymes and metabolic reactions peculiar to only humans were observed. Moreover, when several drugs were exposed to spheroids and hepatotoxicity was evaluated, stable values were obtained for the 50% inhibitory concentration (IC<sub>50</sub>) of albumin secretion at 14 and 21 days. The IC<sub>50</sub> values of most of the tested drugs were lower than in conventional assays, suggesting that the reported evaluation methods might underestimate hepatotoxicity. Furthermore, examination of mRNA expression level and activity of various cytochrome P450 (CYP) after exposure of typical inducers of CYPs to hepatocyte spheroid resulted in a significant increase in the expression level and activity of each. From these results, it was shown that this 3D hepatocyte spheroid system is suitable for follow-up of metabolic processes, long-term tests of hepatotoxicity and enzyme activity induction potential of drugs.</p>

Journal

  • Folia Pharmacologica Japonica

    Folia Pharmacologica Japonica 153(5), 235-241, 2019

    The Japanese Pharmacological Society

Codes

  • NII Article ID (NAID)
    130007649348
  • NII NACSIS-CAT ID (NCID)
    AN00198335
  • Text Lang
    JPN
  • ISSN
    0015-5691
  • NDL Article ID
    029715059
  • NDL Call No.
    Z19-247
  • Data Source
    NDL  J-STAGE 
Page Top