Cd4 isolation kit

Human T Cell Isolation and Culture Leukoreduction chambers from healthy, anonymous donors were purchased from Blood Centers of the Pacific and processed within 12 hours. Primary CD4+ T-cells were harvested by positive selection using a FABian automated enrichment system and CD4 isolation kit (IBA Lifesciences, Goettingen, Germany). Isolated CD4+ T cells were suspended in complete Roswell Park Memorial Institute (RPMI) media, consisting of RPMI-1640 [UCSF Cell Culture Facility (CCF)] supplemented with 5mM 4-(2- hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, UCSF CCF), 2mM Glutamine (UCSF CCF), 50μg/mL penicillin/streptomycin (P/S, UCSF CCF), 5mM nonessential amino acids (UCSF CCF), 5mM sodium pyruvate (UCSF CCF), and 10% fetal bovine serum (FBS, Atlanta Biologicals). These cells were immediately stimulated on anti-CD3 coated plates [coated overnight with 10μg/mL αCD3 (UCHT1, Tonbo Biosciences)] in the presence of 5μg/mL soluble anti-CD28 (CD28.2, Tonbo Biosciences). Cells were stimulated for 48 hours prior to electroporation.

Human T cell isolation and leukoreduction chambers from healthy, anonymous donors were purchased from Blood Centers of the Pacific and processed within 12 h. Primary CD4+ T cells were harvested by positive selection using a FABian automated enrichment system and CD4 isolation kit (IBA Lifesciences). Isolated T cells were suspended in complete RPMI-1640 medium supplemented with 5 mM HEPES, 2 mM glutamine, 50 μg/mL penicillin/streptomycin, 5 mM nonessential amino acids, 5 mM sodium pyruvate, and 10% fetal bovine serum (Atlanta Biologicals). These cells were immediately stimulated on anti-CD3-coated plates (coated overnight with 10 μg/mL anti-CD3 from Tonbo Biosciences) in the presence of 5 μg/mL soluble anti-CD28 (CD28.2, Tonbo Biosciences). Cells were stimulated for 48 h prior to infection.