Fundamental Toxicological Sciences

Paper Details

Fundamental Toxicological Sciences
Vol. 9 No. 4 September 07, 2022 p.135-144
Original Article
Cryopreserved human hepatocytes culture optimization on polymethylpentene oxygen permeable membranes for drug screening purposes
  • Mathieu Anoy (Department of Chemical Engineering, Faculty of Engineering, University of Tokyo /
Mathieu Anoy 1) , Benedikt Scheidecker 1) 2) , Hiroshi Arakawa 3) , Katsuhiro Esashika 4) , Naoki Ishida 3) , Hiroyasu Ito 1) , Hisaaki Yanai 5) , Jun Takahashi 5) , Masaki Nishikawa 1) , Yukio Kato 3) , Yasuyuki Sakai 1)
1) Department of Chemical Engineering, Faculty of Engineering, University of Tokyo , 2) CNRS UMI 2820, Laboratory for Integrated Micro Mechatronic Systems, Institute of Industrial Science, University of Tokyo , 3) Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University , 4) New Business Solutions Department, Innovative Solutions Center for Information & Communication Technology, R&D Center, Mitsui Chemicals, Inc. , 5) Bio Technology & Medical Materials Department, Synthetic Chemicals Laboratory, R&D Center, Mitsui Chemicals, Inc.
Keywords: Cryopreserved hepatocytes, PMP, Oxygen permeable membrane, Drug development

In vitro culture of primary hepatocytes for drug screening purposes remains a challenge as cells rapidly lose their function in conventional culture conditions. Thin oxygen permeable membranes have shown, through direct oxygenation, beneficial effects for long-term culture and cellular function with freshly isolated hepatocytes. However, culture of cryopreserved hepatocytes, a standard for the industry, has shown limits due to high cellular damage, leading to low cellular function. In addition, high sorption of drug screening compounds on PDMS oxygen permeable membranes has rendered evaluation of different molecules, aimed at the improvement of the culture of those cells difficult. Here, culture of cryopreserved hepatocytes was performed on PMP membranes, known to exhibit exceptionally low sorption characteristics. A mixture of anti-apoptotic and anti-inflammatory compounds to improve cell viability during adhesion was tested and evaluation in terms of cellular damage and drug metabolism was performed after 24 hr and 72 hr. Components of the mixture were shown to have beneficial effect on Reactive Oxygen Species production after 6 hr of adhesion as well as on mitochondrial activity and LDH release after 24 hr. Effects in improving recovery of albumin and drug metabolism, could be efficiently measured after 72 hr as a result of the use of PMP. The presented results demonstrate the compatibility of PMP oxygen-permeable membrane-based culture with cryopreserved hepatocytes for efficient drug screening.