A375 atcc crl 1619

The human cutaneous squamous carcinoma cell line SCL-1 was a gift from Prof. Dr. Norbert Fusenig from the German Cancer Research Center (DKFZ, Heidelberg, Germany) (Boukamp et al. 1982 (link)). The human melanoma cell line A375 (ATCC CRL-1619) was obtained from the American Type Culture Collection (ATCC, Virginia, USA). Normal human epidermal keratinocytes (NHEK, C-12001) were ordered from PromoCell (Heidelberg, Germany). SCL-1 and A375 tumor cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, low glucose), supplemented with 10% fetal bovine serum (FBS), streptomycin (100 µg/ml), penicillin (100 U/ml), and GlutaMAX™ (2 mM) at 37 °C in 5% CO₂. NHEK were cultured in keratinocyte growth medium (C-20011, PromoCell) including SupplementMix (C-39016), streptomycin (100 µg/ml) and penicillin (100 U/ml) at 37 °C in 5% CO₂. Subconfluent cells (70–80% confluence) were used for all experiments, if not otherwise stated. For treatment, SCL-1 and A375 were incubated in high glucose (4500 mg/L) DMEM without FBS, while NHEK were grown in keratinocyte basal medium (C-20211, PromoCell). Gossypol and other substances such as ABT-199, inhibitors, and fluorescent dyes were directly added to the cells at the appropriate concentrations.

Human melanoma cells A375 (ATCC^® CRL-1619, from primary cutaneous melanoma) and MeWo (ATCC^® HTB-65, from lymph node cutaneous melanoma metastasis) were obtained from the American Type Culture Collection (ATCC). 501Mel (from cutaneous melanoma metastasis) were kindly provided by Dr. Poliseno (Oncogenomics Unit, Core Research Laboratory, Istituto Toscano Tumori c/o IFC-CNR, Pisa, Italy).
All cell lines were cultured in RPMI 1640 medium (Euroclone, Milan, Italy) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin (Euroclone, Euroclone, Milan, Italy) in a humidified atmosphere containing 5% CO₂ at 37 °C. Cell morphology was examined under light microscopy.

Rutin (Rut), sorbitan monostearate (Span 60) and cholesterol were purchased from Sigma-Aldrich, Merck KgaA, soy lecithin (Lipoid S 75) from Lipoid GmbH (Ludwigshafen, Germany) and absolute ethanol from Merck (Darmstadt, Germany).
The human melanoma cell line—A375 (ATCC^® CRL-1619™) was acquired from American Type Culture Collection (ATCC) and the immortalized human keratinocytes—HaCaT (HaCaT-300493) from CLS Cell Lines Service GmbH. The specific cell culture medium—Dulbecco’s modified Eagle’s medium-high glucose (DMEM), penicillin–streptomycin mixture, phosphate saline buffer (PBS), trypsin-EDTA, Trypan blue, dimethyl sulfoxide (DMSO), and alamarBlue (resazurin) were provided by Sigma-Aldrich, Merck KgaA. The fetal calf serum (FCS) and Hoechst 33342 reagent (Invitrogen) were acquired from Thermo Fisher Scientific, Inc., Waltham, MA, USA.