GATA1 deletion in human pluripotent stem cells increases differentiation yield and maturity of neutrophils


Human pluripotent stem cell (hPSC)-derived tissues can be used to model diseases in cell types that are challenging to harvest and study at-scale, such as neutrophils. Neutrophil dysregulation, specifically neutrophil extracellular trap (NET) formation, plays a critical role in the prognosis and progression of multiple diseases, including COVID-19. While hPSCs can generate limitless neutrophils (iNeutrophils) to study these processes, current differentiation protocols generate heterogeneous cultures of granulocytes and precursors. Here, we describe a method to improve iNeutrophil differentiations through the deletion of GATA1. GATA1 knockout (KO) iNeutrophils are nearly identical to primary neutrophils in form and function. Unlike wild-type iNeutrophils, GATA1 KO iNeutrophils generate NETs in response to the physiologic stimulant lipopolysaccharide, suggesting they are a more accurate model when performing NET inhibitor screens. Furthermore, through deletion of CYBB, we demonstrate that GATA1 KO iNeutrophils are a powerful tool in determining involvement of a given protein in NET formation. © 2023 The Authors.

Authors Harper TC, Oberlick EM, Smith TJ, Nunes DE, Bray MA, Park S, Driscoll CD, Mowbray SF, Antczak C
Journal iScience
Publication Date 2023 Oct 20;26(10):107804
PubMed 37720099
PubMed Central PMC10500457
DOI 10.1016/j.isci.2023.107804

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