Generation of Macrophages from Cynomolgus-Monkey Bone Marrow as a Model to Evaluate Effects of Drugs on Innate Immunity

Nianyu Li, Susan A. Ludmann, Lisa Anest, Ching He, Padma Kumar Narayanan
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Abstract

Macrophages are innate immune cells that play important roles in various physiological and pathological processes. Evaluation of pro-inflammatory effects of drugs on macrophages has become commonplace in preclinical drug development prior to human clinical trials. Despite their body-wide distribution, tissue macrophages are often difficult to collect from large animals and humans in a noninvasive manner. Therefore, in vitro–differentiated macrophages are important tools to facilitate cross-species analysis of macrophage function. Although cynomolgus monkeys are an essential non-rodent species for preclinical research, in vitro differentiation of cynomolgus-monkey macrophages has been poorly characterized. In the present unit, we describe a protocol to differentiate cynomolgus-monkey macrophages from isolated bone marrow mononuclear cells (BMMCs). In contrast to monocytes, cynomolgus-monkey BMMCs show robust expansion in the presence of macrophage colony–stimulating factor in vitro, which allows expansion of many cells from a single animal donor. Macrophages differentiated from BMMCs retain many of the macrophage phenotypes and functions, including phagocytosis and cytokine release, and therefore can be used as a surrogate to assess effects of drugs on cynomolgus-monkey macrophages. © 2019 by John Wiley & Sons, Inc.

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以食蟹猴骨髓巨噬细胞生成为模型评价药物对先天免疫的影响
巨噬细胞是先天免疫细胞,在各种生理和病理过程中发挥重要作用。在人类临床试验之前,评估药物对巨噬细胞的促炎作用已经成为临床前药物开发的常态。尽管组织巨噬细胞遍布全身,但通常很难以无创方式从大型动物和人类身上收集到组织巨噬细胞。因此,体外分化巨噬细胞是促进跨物种巨噬细胞功能分析的重要工具。尽管食蟹猴是临床前研究必不可少的非啮齿类动物,但对食蟹猴巨噬细胞的体外分化却知之甚少。在本单元中,我们描述了一种从分离的骨髓单核细胞(BMMCs)中分化食蟹猴巨噬细胞的方法。与单核细胞相比,在体外巨噬细胞集落刺激因子的存在下,食蟹猴BMMCs表现出强劲的扩张,这使得单个动物供体可以扩增许多细胞。从BMMCs分化出来的巨噬细胞保留了许多巨噬细胞的表型和功能,包括吞噬和细胞因子释放,因此可以作为评估药物对食蟹猴巨噬细胞影响的替代品。©2019 by John Wiley &儿子,Inc。
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