Rpmi 1640 culture medium

The human monocytic leukemia cell line THP-1 (American Type Culture Collection—ATCC, Manassas, VA, USA) was grown in RPMI 1640 culture medium (ATCC) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin both from Thermo Fisher Scientific (Waltham, MA, USA). Cells were cultured at 37 °C in 5% CO₂ in a humidified incubator. Media were renewed every 3 days and cells were sub-cultured when the concentration reached 8 × 10⁵ cells/mL. Cells were discarded and replaced by frozen stocks after 20 passages. Cytotoxicity kit for determining cell viability based on lactate dehydrogenase (LDH) release was purchased from Thermo Fisher Scientific. Lipopolysaccharide (LPS) was purchased from Millipore-Sigma (St. Louis, MO, USA). ELISA kits were purchased from Thermo Fisher Scientific (Waltham, MA, USA).

The three cell lines used were all cultured in conformity with the sterile technique and the standard mammalian cell culture protocols under a 5% CO₂ atmosphere at 37 °C.
Lymphocyte cell line (IST-EBV-TW6B) was purchased from the cell bank IRCCS AOU San Martino IST (Italy). Cells were cultured in advanced RPMI 1640 culture medium (Gibco) with 20% of heat inactivated fetal bovine serum (FBS, Gibco), 1% penicillin/streptomycin (P/S, Sigma) and 1% of l-Glutamine 200 mM (Lonza) in 75 cm² not treated cell culture flasks (Corning) maintaining the cell density between 9 × 10^4–5 cells/mL.
Daudi cells (ATCC® CCL­213™), originating from a Burkitt’s lymphoma patient, were obtained from American Type Culture Collection (ATCC). Cells were grown in RPMI 1640 culture medium (ATCC) supplemented with 10% of heat inactivated FBS (ATCC), 1% P/S (Sigma) in 75 cm² not treated cell culture flasks (Corning) with a cell density between 3 × 10^5–6 cells/mL.
HL60 cells (ATCC® CCL-240™), from an acute myeloid leukemia patient, were purchased from ATCC. They were maintained in Iscove’s Modified Dulbecco’s Medium (Sigma) with 20% heat inactivated FBS (Sigma), 1% Glutamine (Sigma), 1% P/S (Sigma) in 75 cm² not treated cell culture flasks (Corning), adjusting cell density to 1 × 10^5–6 cells/mL.

All cell lines were cultured according to standard mammalian cell culture protocols and a sterile technique at 37 °C under a 5% CO₂ atmosphere.
Daudi cells (ATCC^® CCL-213™), derived from a patient affected by Burkitt’s lymphoma, were obtained from American Type Culture Collection (ATCC). Cells were cultured in RPMI 1640 culture medium (ATCC) supplemented with 10% of heat-inactivated fetal bovine serum (FBS, ATCC), 1% penicillin/streptomycin (P/S, Sigma) in 75 cm² non-treated cell culture flasks (Corning). The cell density was maintained of 3 × 10^5–6 cells/mL.
The lymphocyte cell line (IST-EBV-TW6B) was purchased from the cell bank IRCCS AOU San Martino IST (Italy). Cells were grown in advanced RPMI 1640 culture medium (Gibco) with 20% heat-inactivated FBS (Gibco), 1% L-glutamine 200 mM (Lonza), and 1% P/S (Sigma) in 75 cm² non-treated cell culture flasks (Corning) with a cell density of 9 × 10^4–5 cells/mL.
HL60 cells (ATCC^® CCL-240™), derived from an acute myeloid leukemia patient, were obtained from ATCC. They were maintained in Iscove’s Modified Dulbecco’s Medium (Sigma) with 20% heat-inactivated FBS (Sigma), 1% glutamine (Sigma), 1% P/S (Sigma) in 75 cm² non-treated cell culture flasks (Corning), adjusting the cell density to 1 × 10⁵⁻⁶ cells/mL.