Rpmi 1640

Human dermal fibroblasts from healthy adult donors (aged 35–45) were obtained from the “Cell Line and DNA Biobank from Patients Affected by Genetic Diseases—NETWORK OF GENETIC BIOBANKS TELETHON” [43 ]. Fibroblasts were used for the experiments between passages 11 and 13. The experiments were performed using fibroblasts from two different donors.
Fibroblasts were grown in RPMI-1640 (Carlo Erba Reagents, Milan, Italy) supplemented with 10% Fetal Bovine Serum (FBS, Carlo Erba Reagents, Milan, Italy) and 2 mM L-Glutamine (Carlo Erba Reagents, Milan, Italy). Cultures were incubated at 37 °C in 95% humidity and 5% CO₂ atmosphere. At confluence, fibroblasts were detached from culture flasks by treatment with trypsin (Carlo Erba Reagents, Milan, Italy). Cells were thawed on an average of two weeks before each experiment to have sufficient numbers of cells. Cells were routinely checked for the presence of mycoplasma by using the nested PCR method described in Tang et al. [44 (link)].

Human cutaneous melanoma cell lines (Mel 195, 275, 313, 346, 116, 120, 514, 142, 237, 403, 458, 345, 599, and 261) were generated from surgically removed metastatic lesions from melanoma patients, as previously described (Altomonte et al., 1993 (link)). Human hematological cancer cell lines (Daudi, HL-60, NALM-6, Raji, U-937, KG-1a, Jurkat, JY, Ri-1, K562) were purchased from American Type Culture Collection (Rockville, MD, United States).
Melanoma cells were grown in RPMI 1640 (Carlo Erba, Milan, Italy) supplemented with 10% heat-inactivated FBS (Biochrom, Berlin, Germany) and 2 mM L-glutamine (Biochrom, Berlin, Germany). Hematological tumor cell lines were grown in ISCOVE Basal Medium (Biochrom, Berlin, Germany) supplemented with 10% heat-inactivated FBS, 2 mM L-glutamine and 100 μg/μl penicillin/streptomycin (Biochrom, Berlin, Germany).