Division of Genetic and Molecular Toxicology, National Center for Toxicological Research, Jefferson, Arkansas.
Toxicologic Pathology Associates, Jefferson, Arkansas.
Curr Protoc. 2024 Sep;4(9):e70015. doi: 10.1002/cpz1.70015.
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.
原代人肝细胞 (PHHs) 被认为是体外评估各种药物或化学物质毒性的“金标准”。然而，由于其可用性有限，从啮齿动物中分离的原代肝细胞在各种实验研究中比 PHHs 更常用。然而，与人类和非人类灵长类动物相比，人类和大鼠在药物代谢方面的差异更大。在这里，我们描述了一种从恒河猴 (Macaca mulatta，一种旧世界猴) 肝脏原位全肝灌流后分离原代肝细胞的方法。还描述了冷冻保存和恢复原代猕猴肝细胞 (PMHs) 的技术。鉴于非人类灵长类动物与人类在生理和遗传上的惊人相似性，使用该方案分离的 PMHs 可作为毒理学研究和临床前研究中 PHHs 的可靠替代物。发表于 2024 年。本文是美国政府的工作，在美国属于公有领域。 基本方案 1：原位全肝灌流 基本方案 2：原代猕猴肝细胞分离和细胞铺板 基本方案 3：原代猕猴肝细胞的冷冻保存和恢复
