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. 2021 Jan 22;11:602530. doi: 10.3389/fimmu.2020.602530

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Copyright © 2021 Yang, Lode, Berzaghi, Islam, Martinez-Zubiaurre and Hellevik

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Natural killer (NK) cell purity, cancer-associated fibroblast (CAF) characterization, and radiation-induced morphological changes in CAFs. (A) Dot plots from flow cytometer analyses, illustrating 93% purity of NK cells defined as CD56⁺CD3⁻ upon negative selection by antibody-coated microbeads. (B) Immunostaining of cultivated CAFs, using CAF-specific FAP-1 antibody (red) and nuclear DAPI (blue). (C) Flow cytometry analyses of CAFs after immunostaining with CAF-specific αSMA antibody. Negative control is CAFs stained with isotype control antibody. (D) Representative culture of spindle-shaped, non-irradiated human lung CAFs at passage three. (E) Representative image of CAFs acquired 5 days after exposure to ionizing radiation (1x18 Gy). (F) Image of irradiated CAFs in culture, acquired 2 days after third dose of 6 Gy (3x6 Gy). (G) Culture of normal skin fibroblasts (NFs). (H) Image of non-adherent (CD56⁺CD3⁻) NK cells in monoculture. (I) Image of co-cultures consisting of adherent CAFs and non-adherent NK cells.