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OMIP ‐070: NKp46 ‐Based 27‐Color Phenotyping to Define Natural Killer Cells Isolated From Human Tumor Tissues
Author(s) -
Frutoso Marie,
Mair Florian,
Prlic Martin
Publication year - 2020
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.24230
Subject(s) - collagenase , cytometry , flow cytometry , biology , cell , immunology , microbiology and biotechnology , chemistry , biochemistry , enzyme
Purpose and appropriate samples 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.