Effects of High Concentration Potassium Chloride on the Preparation and Immunophenotype of CAR-T Cells
WANG Yao
GAO Yao
HE Meijie
LI Linlin
CHEN Guangyu
XU Benling
LI Tiepeng
Abstract：Objective To investigate the effects of high concentration potassium chloride on the in vitro expansion,transduction efficiency,memory phenotype,and cytotoxicity of CD19-specific chimeric antigen receptor(CAR)T cells.Methods Density gradient centrifugation was used to isolate human peripheral blood mononuclear cells,and CD3 MicroBeads were introduced to positively sort T cells.T cells were activated with anti-CD3 monoclonal antibody and RetroNectin followed by transducing with a lentiviral vector expressing CD19-specific CAR.Subsequently,T cells were divided into the control and the high concentration potassium chloride group.The trypan blue staining was used to count the number of T cells.Flow cytometer was used to analyze the transduction efficiency,the proportions of CD4+T,CD8+T,and CD3+effector memory T cells(TEM)and the cytotoxicity against CD19-positive OCI-Ly8 cells.Results Compared with the control group,the high concentration of potassium chloride did not affect the in vitro expansion of the T cells,the transduction efficiency,the proportions of CD4+T cells,the proportions of CD8+T cells and the cytotoxicity of CAR-T cells(P＞0.05).However,the high concentration of potassium chloride significantly increased the proportions of CD3+TEM cells(P＜0.05).Conclusion The high concentration of potassium chloride can be used to optimize the culture of CAR-T cells in vitro by increasing the proportions of CD3+TEM cells.
Keywords：potassium chloridechimeric antigen receptoreffector memory T cells
Publication Date：2025-12-13
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：4( 4225-4228 )