Neutrophil isolation from nonhuman species.
Neutrophil isolation from nonhuman species.
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DOI：
10.1007/978-1-62703-845-4_3
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Quinn MT
中科院分区：
文献类型：
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作者：
Siemsen DW;Malachowa N;Schepetkin IA;Whitney AR;Kirpotina LN;Lei B;Deleo FR;Quinn MT
中性粒细胞生物化学理解的新进展需要有效的程序来分离纯化的中性粒细胞群体。尽管人类中性粒细胞分离的方法现已成为标准，但从用于模拟人类疾病的许多非人类物种中分离中性粒细胞的类似程序尚未得到很好的开发。由于中性粒细胞是反应性细胞，因此分离方法对于避免分离技术引起的细胞功能改变极其重要。我们在此介绍了从大型动物（牛、马、羊）、小动物（鼠和兔）和非人灵长类动物（食蟹猴）中可重复地分离高度纯化的中性粒细胞的方法，并描述了获得最高细胞纯度、产量和活力的优化细节。我们还描述了使用基于流式细胞术的吞噬测定来验证纯化细胞群中吞噬能力的方法。
The development of new advances in the understanding of neutrophil biochemistry requires effective procedures for isolating purified neutrophil populations. Although methods for human neutrophil isolation are now standard, similar procedures for isolating neutrophils from many of the nonhuman species used to model human diseases are not as well developed. Since neutrophils are reactive cells, the method of isolation is extremely important to avoid isolation technique-induced alterations in cell function. We present methods here for reproducibly isolating highly purified neutrophils from large animals (bovine, equine, ovine), small animals (murine and rabbit), and nonhuman primates (cynomolgus macaques), and describe optimized details for obtaining the highest cell purity, yield, and viability. We also describe methods to verify phagocytic capacity in the purified cell populations using a flow cytometry-based phagocytosis assay.