This 27‐color panel has been validated and optimized to comprehensively profile natural killer (NK) cells isolated from human tumors using a collagenase Type II‐based digestion protocol. We confirmed that detection of protein expression by antibodies used in our final panel was not affected during tissue digestion. During this evaluation process, we found that detection of CD56, a biomarker typically used to identify NK cells, was affected substantially by collagenase‐based digestion. Thus, our panel is centered around expression of NKp46, which is sufficient to identify NK cells and not affected by the tissue collagenase digestion step. Our panel further includes biomarkers used to extrapolate NK‐cell maturation, differentiation, migration, homing potential, and functional state. Our panel is intended to provide in‐depth characterization of human NK cells isolated from tissues, which we specifically tested using oral squamous cell carcinomas tissues, but it is compatible with other tissues that can be dissociated with a collagenase Type II‐based protocol. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.

中文翻译：

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OMIP-070：基于 NKp46 的 27 色表型分析以定义从人类肿瘤组织中分离的自然杀伤细胞

这个 27 色面板已经过验证和优化，可以使用基于 II 型胶原酶的消化方案全面分析从人类肿瘤中分离的自然杀伤 (NK) 细胞。我们证实，在组织消化过程中，我们最终面板中使用的抗体对蛋白质表达的检测没有受到影响。在此评估过程中，我们发现通常用于识别 NK 细胞的生物标志物 CD56 的检测受到基于胶原酶的消化的显着影响。因此，我们的面板以 NKp46 的表达为中心，这足以识别 NK 细胞并且不受组织胶原酶消化步骤的影响。我们的面板还包括用于推断 NK 细胞成熟、分化、迁移、归巢潜力和功能状态的生物标志物。我们的面板旨在提供从组织中分离的人类 NK 细胞的深入表征，我们使用口腔鳞状细胞癌组织专门测试了这些细胞，但它与可以使用基于 II 型胶原酶的方案解离的其他组织兼容。© 2020 作者。细胞计数 A 部分由 Wiley Periodicals LLC 代表国际细胞计数促进协会出版。