Comparative Study of Culture Conditions for Maintaining CYP3A4 and ATP-Binding Cassette Transporters Activity in Primary Cultured Human Hepatocytes

Journal of Pharmacological Sciences

Hiroshi Nakano²  Toshio Kumai³  Yuko Takeba¹  Sachiko Takenoshita-Nakaya¹  Naoki Matsumoto¹  Yoshie Harimoto¹  Yuichi Kinoshita¹  Shinichi Kobayashi¹  Takehito Ohtsubo² 

[1] Department of Pharmacology, St. Marianna University School of Medicine, Japan;Division of Gastroenterological Surgery, St. Marianna University School of Medicine, Japan;Department of Pharmacogenomics, St. Marianna University School of Medicine, Japan

关键词: CYP3A4;    HNF-4α;    primary hepatocyte;    ATP-binding cassette transporter;    Lanford’s medium;   

DOI  :  10.1254/jphs.10215FP

学科分类：药学

来源: Nihon Yakuri Gakkai Henshuubu / Japanese Pharmacological Society

【 摘 要 】

References(39)Cited-By(6)The aim of this study was to determine suitable culture conditions for maintaining the activity of cytochrome p450 (CYP) 3A4 and drug transporters in primary cultured human hepatocytes. Human hepatocytes were isolated using the two-step collagenase perfusion technique and were cultured with four different media, serum-free William’s E medium (serum-free WEM), WEM containing fetal calf serum (FCS-WEM), WEM with human serum (HS-WEM), and Lanford’s medium. The albumin levels were maintained for 7 days in hepatocytes. Although CYP3A4 mRNA levels gradually decreased from 3 days, CYP3A4 and hepatocyte nuclear factor-4α alpha protein levels and activities were maintained for 7 days in hepatocytes cultured with serum-free WEM and Lanford’s but not in those with FCS-WEM and HS-WEM. Furthermore, CYP3A4 protein levels were significantly increased by the addition of rifampicin and dexamethasone to the culture media, indicating that the induction potential was maintained. The protein levels of P-glycoprotein, multi-drug-resistance-2, and breast cancer-resistance protein were maintained for 7 days in all media. Serum-free WEM and Lanford’s also maintained protein levels of CYP2C19, CYP2D6, and organic anion transporter polypeptide in the hepatocytes. Serum-free WEM and Lanford’s may be appropriate culture media for maintaining CYP3A4 and drug transporter protein levels in primary cultured hepatocytes.

【 授权许可】

Unknown