An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - are your sorted cells getting SICS?

Pfister, G; Toor, SM; Sasidharan Nair, V; Elkord, E

Abstract

Flow cytometry and fluorescence-activated cell sorting have become invaluable tools to analyze and isolate specific cell populations in a wide range of biomedical research and clinical applications. In countless approaches worldwide, scientists are using single cell analyses to better understand the significance and variation within different cellular populations, and fluorescence-activated cell sorting has become a major technique for cell isolation in both basic and clinical research. However, majority of available cell sorters are pressurized, droplet-based systems, which apply significant environmental pressure and shear stress to cells during sorting. Recently, the flow cytometry community has become increasingly aware about the potential negative effects this could have on sorted cells and the term "sorter induced cell stress" (SICS) has been proposed. However, up to date only a limited number of studies have investigated the effects of cell sorting on cell viability and function. Therefore, solid data on the effects of sheath pressure and nozzle size on survival and function of sorted cells are surprisingly rare. With this in mind, we sorted "CD4 " T-cells and "live" cells from human peripheral blood mononuclear cells (PBMCs) at different sort conditions and analyzed their quality before and after sorting in a series of assays. Here we present our findings in reference to cell viability and cell proliferation following sorting on different instruments (BD FACSAria III SORP and BD FACSJazz), utilizing different nozzle sizes (70 to 100 μm) and sheath pressure settings (20 to 70 psi). The results show no significant differences in cell viability and proliferation after the different tested sort conditions, but rather differences between individual experiments. These findings are evaluated and their potential significance in cell sorting experiments is discussed. [Abstract copyright: Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.]

Citation

Pfister, G., Toor, S., Sasidharan Nair, V., & Elkord, E. (2020). An evaluation of sorter induced cell stress (SICS) on peripheral blood mononuclear cells (PBMCs) after different sort conditions - are your sorted cells getting SICS?. Journal of Immunological Methods, 487, 112902. https://doi.org/10.1016/j.jim.2020.112902

• Journal Article Type: Article
• Acceptance Date: Oct 13, 2020
• Online Publication Date: Oct 15, 2020
• Publication Date: Dec 1, 2020
• Deposit Date: Nov 3, 2020
• Publicly Available Date: Nov 3, 2020
• Journal: Journal of Immunological Methods
• Print ISSN: 0022-1759
• Publisher: Elsevier
• Volume: 487
• Pages: 112902
• DOI: https://doi.org/10.1016/j.jim.2020.112902
• Publisher URL: https://doi.org/10.1016/j.jim.2020.112902
• Related Public URLs: http://www.journals.elsevier.com/journal-of-immunological-methods/
• Additional Information: Additional Information : ** From PubMed via Jisc Publications Router **Journal IDs: eissn 1872-7905 **Article IDs: pubmed: 33069766; pii: S0022-1759(20)30196-4 **History: accepted 13-10-2020; revised 10-09-2020; submitted 18-06-2020; Funders : Qatar Biomedical Research Institute, Qatar Foundation;Qatar National Library; Grant Number: VR04