Development of a High-Color Flow Cytometry Panel for Immunologic Analysis of Tissue Injury and Reconstruction in a Rat Model.

Autor:	Adusei KM; Section on Immunoengineering, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA.; Biomedical Engineering and Technology Acceleration (BETA) Center, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA., Ngo TB; Section on Immunoengineering, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA.; Biomedical Engineering and Technology Acceleration (BETA) Center, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA., Alfonso AL; Section on Immunoengineering, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA.; Biomedical Engineering and Technology Acceleration (BETA) Center, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA., Lokwani R; Section on Immunoengineering, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA.; Biomedical Engineering and Technology Acceleration (BETA) Center, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA., DeStefano S; Section on Immunoengineering, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA.; Biomedical Engineering and Technology Acceleration (BETA) Center, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA., Karkanitsa M; Section on Immunoengineering, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA.; Biomedical Engineering and Technology Acceleration (BETA) Center, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA., Spathies J; Biomedical Engineering and Technology Acceleration (BETA) Center, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA.; Bioengineering and Physical Sciences Shared Resource, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA., Goldman SM; DoD-VA Extremity Trauma & Amputation Center of Excellence, Bethesda, Maryland, USA.; Department of Surgery, Uniformed Services University of the Health Sciences, Walter Reed National Military Medical Center, Bethesda, Maryland, USA., Dearth CL; DoD-VA Extremity Trauma & Amputation Center of Excellence, Bethesda, Maryland, USA.; Department of Surgery, Uniformed Services University of the Health Sciences, Walter Reed National Military Medical Center, Bethesda, Maryland, USA., Sadtler KN; Section on Immunoengineering, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA.; Biomedical Engineering and Technology Acceleration (BETA) Center, National Institute for Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA.
Jazyk:	angličtina
Zdroj:	Cells, tissues, organs [Cells Tissues Organs] 2023; Vol. 212 (1), pp. 84-95. Date of Electronic Publication: 2022 Apr 22.
DOI:	10.1159/000524682
Abstrakt:	The rat model is an important resource in biomedical research due to its similarities to the human immune system and its use for functional studies. However, because of the preponderance of mouse models in foundational and mechanistic immunological studies, there is a relative lack of diverse, commercially available flow cytometry antibodies for immunological profiling in the rat model. Available antibodies are often conjugated to common fluorophores with similar peak emission wavelengths, making them hard to distinguish on conventional flow cytometers and restricting more comprehensive immune analysis. This can become a limitation when designing immunological studies in rat injury models to investigate the immune response to tissue injury. In addition, this lack of available antibodies limits the number of studies that can be done on the immune populations in lymphoid organs in other research areas. To address this critical unmet need, we designed a spectral flow cytometry panel for rat models. Spectral cytometry distinguishes between different fluorophores by capturing their full emission spectra instead of their peak emission wavelengths. This flow cytometry panel includes 24 distinct immune cell markers to analyze the innate and adaptive immune response. Importantly, this panel identifies different immune phenotypes, including tolerogenic, Type 1, and Type 2 immune responses. We show that this panel can identify unique immune populations and phenotypes in a rat muscle trauma model. We further validated that the panel can identify distinct adaptive and innate immune populations and their unique phenotypes in lymphoid organs. This panel expands the scope of previous rat panels providing a tool for scientists to examine the immune system in homeostasis and injury while pairing mechanistic immunological studies with functional studies. (© 2022 S. Karger AG, Basel.)
Databáze:	MEDLINE
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