Isolation and Cryopreservation of Primary Macaque Hepatocytes

Abstract Primary human hepatocytes (PHHs) are recognized as the "gold standard" for evaluating toxicity of various drugs or chemicals in vitro. However, due to their limited availability, primary hepatocytes isolated from rodents are more commonly used in various experimental studies than PHHs. However, bigger differences in drug metabolism were seen between humans and rats compared to those between human and non‐human primates. Here, we describe a method to isolate primary hepatocytes from the liver of rhesus macaques ( Macaca mulatta , a species of Old‐World monkey) after in situ whole liver perfusion. Techniques for cryopreserving and recovering primary macaque hepatocytes (PMHs) are also described. Given the remarkable physiological and genetic similarity of non‐human primates to humans, PMHs isolated using this protocol may serve as a reliable surrogate of PHHs in toxicological research and preclinical studies. Published 2024. This article is a U.S. Government work and is in the public domain in the USA. Basic Protocol 1 : In situ whole liver perfusion Basic Protocol 2 : Primary macaque hepatocyte isolation and cell plating Basic Protocol 3 : Cryopreservation and recovery of primary macaque hepatocytes