Jurkat T Cell Culture

Jurkat T cells, clone E6-1 (ATCC: TIB-152), were cultured in a humidified incubator at 37°C under 5% CO₂ in RPMI 1640 medium (VWR) supplemented with 10% fetal bovine serum (FBS) and 1% antibiotics. Medium was replaced every 2–3 days and cell density was maintained below 1 × 10⁶ cells/mL for cell propagation. Three types of plates were examined for suspension cell confinement in the PEG-BSA system: flat-bottom, round-bottom and V-bottom. Jurkat T cells were confined in the BSA phase (bottom). The BSA-phase was labeled with FITC-conjugated Dex and the cells were labeled with CellTracker Red™ CMTPX (Life Technologies) to aid in visualization. Cells cultured in the PEG-BSA system were observed using a 4x objective lens on a Nikon Eclipse Ti microscope.