Hut78

Hepatoma cell lines FLC-4 were provided by Dr S. Nagamori (Kyorin University, Japan). Breast adenocarcinoma cell lines, MCF7, and gastric adenocarcinoma cell lines, AGS, were provided by the Department of Chest, Breast and Endocrine Surgery, Akita University, Japan. Colon carcinoma cell lines, HCT116, and T-cell leukemia cell lines, Jurkat, were provided by Dr H. Tomoda (Kitasato University, Japan). Esophageal carcinoma cell lines, TE4, and T-cell leukemia cell lines, HPB-ALL, HUT78 and PEER, were obtained from Riken Bioresource Center, Japan. Embryonic kidney cell lines, HEK293, were provided by the Department of Hematology, Nephrology and Rheumatology, Akita University. Promyelocytic leukemia cell lines, HL-60, and T-cell leukemia cell lines, JKT-beta-del, were obtained from the Japanese Collection of Research Bioresources, Japan. FLC-4, MCF7 and HCT116 cells were cultured in DMEM/F12, Eagle's MEM containing 10 μg/mL insulin and McCoy's 5A, respectively. AGS and HEK293 cells were cultured in DMEM. TE4, Jurkat, HPB-ALL, HUT78, PEER, HL-60 and JKT-beta-del cells were cultured in PRMI-1460. Each medium was supplemented with 10% FBS, 100 units/mL penicillin, 100 μg/mL streptomycin and 2.5 μg/mL amphotericin B. Cells were cultured in a humidified 5% CO₂ atmosphere at 37°C.

As a model of endothelial cells within blood vessels, we used the representative human umbilical vein endothelial cell line EAhy926. The latter was kindly provided by Dr Edgell (University of North Carolina, Chapel Hill, NC, USA). As model of hepatocytes, we used HepG2 cells (Riken BioResource Center (BRC), Ibaraki, Japan). EAhy926 and HepG2 cells were cultured in Dulbecco’s modified Eagle’s medium with low-glucose media (Wako Pure Chemicals, Osaka, Japan) supplemented with 10% fetal bovine serum (Sigma–Aldrich Japan) and 50 U/mL penicillin-streptomycin. HUT78 (CTCL), Molt4 (acute T-lymphoblastic leukemia), and Ramos (Burkitt lymphoma) lymphoid neoplastic cell lines were obtained from Riken BRC and the American Type Culture Collection (Manassas, VA, USA) and cultured in RPMI1640 medium (Wako Pure Chemicals) supplemented with 10% fetal bovine serum and 50 U/mL penicillin-streptomycin. We could not obtain sufficient numbers of vascular endothelial cells and lymphoma cells for assays, so we used these cell lines. To assess the various effects of antineoplastic drugs, those cell lines with each malignant cell type are most popularly used in the world.

Human T cell acute lymphoblastic leukemia cell lines (CCRF-CEM, Jurkat, TALL1) and the human Sezary syndrome cell line (HUT78) were obtained from RIKEN BRC. The canine T cell acute lymphoblastic leukemia cell line (UL-1) was kindly provided by Dr Hajime Tsujimoto. MycoAlert Mycoplasma Detection kit (Lonza) was used to test mycoplasma contamination. The cells were cultured in RPMI1640 (Sigma-Aldrich) containing 10% fetal bovine serum (FBS, Nichirei Biosciences) and antibiotic/antimycotic (Nacalai Tesque).