Converting suspension-type human primary hepatocytes to be plateable-type for use as an in vitro pharmacokinetics model

Abstract

Preclinical animal models cannot precisely predict medication failures in clinical trials. Human primary hepatocytes (HPHs) are the gold standard for in vitro culture systems that recapitulate in vivo human liver characteristics. Consequently, demand for HPHs is rising despite their high cost, batch-to-batch variability, and inadequate supply. Existing commercial HPHs are plateable (pHPHs) and suspension (sHPHs) using same isolation and cryopreservation technique. The sHPHs cannot be cultured on the plate surface or cannot efficiently relocalize efflux transporters. They can also have a life of maximum of 2–4 hours. Therefore, they are unsuitable for assays requiring well-organized efflux transporters or long-term studies, such as enzyme induction or metabolic assays for low-turnover drugs. We aim to develop a novel pharmacokinetic model of converted sHPHs to be attached like pHPHs. We successfully cultured sHPHs as 2D sandwich-cultured sHPHs for use as a promising, reproducible, and affordable in vitro model for efflux trans- porter assays, cytochrome P450 induction, and metabolic studies. The sHPHs successfully attached, converted to be plateable, similar morphology, and mRNA expression as pHPHs, as well as detectable metabolic activity and extended lifespan. Overall, this study is the first to culture functional 2D sandwich sHPHs as a novel in vitro pharmacokinetic model.