Isolation and Cryopreservation of Primary Macaque Hepatocytes

Xiaoqing Guo, Ji-Eun Seo, Kelly Davis, Pritpal Malhi, Cameron Fili
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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

原代猕猴肝细胞的分离与冷冻保存
原代人类肝细胞(PHHs)被公认为体外评估各种药物或化学品毒性的 "黄金标准"。然而,由于从啮齿类动物中分离的原代肝细胞的可用性有限,因此在各种实验研究中,从啮齿类动物中分离的原代肝细胞比从人类肝细胞中分离的原代肝细胞更常用。然而,与人和非人灵长类动物相比,人和大鼠在药物代谢方面的差异更大。在此,我们介绍了一种原位全肝灌注后从猕猴肝脏中分离原代肝细胞的方法。此外,还介绍了冷冻保存和复原原代猕猴肝细胞(PMHs)的技术。鉴于非人类灵长类动物在生理和遗传方面与人类具有明显的相似性,采用该方案分离的原代猕猴肝细胞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。基本方案 1:原位全肝灌注基本方案 2:原代猕猴肝细胞分离和细胞培养基本方案 3:原代猕猴肝细胞的冷冻保存和回收
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