Cell Culture Protocols for Cancer and Immortalized Cell Lines

Rabbit cells (cell line RK13; ATCC CCL-37) and MDA-435 (cell line MDA-MB-435; ATCC HTB-129) and PANC-1 (32 (link)) human cancer cells were cultured in Dulbecco minimum essential medium (DMEM; Invitrogen, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, USA), 2 mM glutamine (Invitrogen, USA), and 100 μg/mL of penicillin-streptomycin (Pen/Strep; Invitrogen, USA). Hare cells (cell line HN-R; Friedrich Loeffler Institut, Germany), a spontaneously immortalized cell line derived from kidney (41 (link)), were cultured in 50% Iscove’s modified Dulbecco’s medium (IMDM) and 50% Ham’s F-12 medium supplemented with 10% FBS, 2 mM glutamine, and 100 μg of Pen/Strep per mL. All cultures were maintained at 37°C in a humidified 5% CO₂ incubator (Thermo Fisher, USA). The RL-5 rabbit CD4⁺ T cell line was cultured in RPMI 1640 (obtained from Gibco), also supplemented with 10% FBS, 2 mM glutamine, and 100 g/mL of Pen/Strep.