Crl 1790

Seven different cell lines, HeLa (ATCC^® CCL-2™), HepG2 (ATCC^® HB-8065™), MCF-7 (ATCC^® HTB-22™), SW-620 (ATCC^® CCL-227™), HCT116 (ATCC^® CCL-247™), CCD 841 CoN (ATCC^® CRL-1790™) and normal human dermal fibroblast (NHDF) (ATCC^® PCS-201-012™) were used in this study. Each cell line was cultured in CO₂ incubator at 37 °C and 5% CO₂. Cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS), 60 mg/mL of penicillin and 100 mg/mL of streptomycin.

This study was carried out with use of three human cell lines: normal colon epithelial cell line CCD 841 CoN (ATCC^® CRL-1790™, Manassas, VA, USA), gastric adenocarcinoma cell line AGS (ATCC^® CRL-1739™, Manassas, VA, USA) and human colorectal adenocarcinoma cell line HT-29 (ATCC^® HTB-38™, Manassas, VA, USA). These cell lines were obtained from the American Type Culture Collections (ATCC, Manassas, VA, USA). Cell cultures were maintained under controlled conditions (temperature 37 °C; atmosphere: air 95%; CO₂ 5%;) in appropriate medium supplemented with 10% FBS according to the ATCC protocol.

The human colorectal adenocarcinoma cell line HT-29 (Cat. no: 85061109), human gastric carcinoma Hs 746T cell line (ATCC^® HTB-135™), human colon cancer Caco-2 cell line (ATCC^® HTB-37™), and human normal colon mucosa CCD 841 CoN cell line (ATCC^® CRL-1790™) were used in this study and cultured according to the method that was previously described in detail in this report [23 (link)]. The cell viability and metabolic activity were determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay [78 (link)]. The first cytotoxic dose (IC₁₀), the median effective concentration (IC₅₀), and the lethal dose (IC₉₀) were calculated on the basis of the MTT results.

E705 and MICOL24 colon cancer cells (kindly provided by Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy) were grown in RPMI 1640 medium supplemented with heat-inactivated 10% fetal bovine serum (FBS), 2 mM l-glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin and were maintained at 37 °C in a humidified 5% CO₂ incubator. The CACO-2 (ATCC^® HTB-37™) colon cancer cell line and CCD 841 (ATCC^® CRL-1790™) healthy mucosa cell line were grown in EMEM medium supplemented with heat-inactivated 10% FBS, 2 mM l-glutamine, 1% nonessential amino acids, 100 U/mL penicillin, 100 μg/mL streptomycin and maintained at 37 °C in a humidified 5% CO₂ incubator. DIFI human CRC cells, kindly provided by Josep Tabernero (Vall d’Hebron Institute of Oncology, Vall d’Hebron University Hospital, Universitat Autònoma de Barcelona, Spain), were grown in Ham’s F12 medium supplemented with heat-inactivated 5% FBS, 2 mM l-glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin and were maintained at 37 °C in a humidified 5% CO₂ incubator. ATCC^® validated cell lines by short tandem repeat profiles that are generated by simultaneous amplification of multiple short tandem repeat loci and amelogenin (for gender identification). All the reagents for cell culture were supplied by Lonza (Lonza Group, Basel, Switzerland).

Colorectal cancer (CRC) cell lines HCT-15 and WiDr were purchased from American Type Culture Collection (ATCC). HCT-15 was cultured in DMEM high glucose (Gibco, Gaithersburg, MD, USA), while WiDr was cultured in MEM (Gibco). All CRC cells were authenticated using STR profiling. A normal human colon epithelial cell line, CRL-1790, also purchased from ATCC, was cultured in Dulbecco’s modified Eagle’s Medium (DMEM) F-12 (Gibco). Human bone marrow-derived mesenchymal stem cells (BM-MSC), obtained from Cryocord Sdn Bhd (Malaysia), was cultured in DMEM F-12. All media were supplemented with 10% fetal bovine serum (FBS) (Gibco) and 1% penicillin–streptomycin (Gibco). The cells were cultured under standard culture conditions at 37 °C in 5% CO₂.

We have previously described iPSC lines derived from two MSCs, namely adipose stem cell (ASC; Invitrogen, Carlsbad, CA, USA) and human adipose-derived MSC (MSC-AT; PromoCell, Heidelberg, Germany), and from a human white pre-adipocyte (HWP) cell line [4 (link), 27 (link)]. In this work, human adipose-derived MSC, designated ASC Lonza, was purchased from Lonza, Lonza, Verviers, Belgium. MH#1 was an iPSC cell lined established from ASC Lonza in our lab (S. Sugii, unpublished data). WJ0706 is a human MSC cell line derived from Wharton’s Jelly (WJ) obtained from Cytopeutics Sdn. Bhd, Selangor, Malaysia (http://www.cytopeutics.com). The MSC cell lines were isolated and characterized at Cytopeutics according to standard procedures and with ethical clearance [28 (link)]. Human placenta choriocarcinoma cell line JEG-3 (ATCC HTB-36), human normal placental cell line HS 799. PI (ATCC CRL-7530) and human normal colon cell line CRL-1790 (ATCC CRL-1790) were purchased from ATCC (Manassas, VA, USA). Cancer cell lines were kindly provided by Professor Y.M. Lim, Cancer Research Center, Universiti Tunku Abdul Rahman.

Two cervical cancer cell lines, HeLa and HT-3 (ATCC, Manassas, VA, USA), and two control cell lines, human epidermal keratinocytes HK (Fisher Scientific, Loughborough, UK) and human colon epithelial cells CRL-1790 (ATCC, Manassas, VA, USA), were employed in this study. HeLa cells (HPV-18-positive) and HT-3 (HPV-negative) were cultured in RPMI 1640 complete media, and CRL-1790 were grown in DMEM media, containing 10% foetal calf serum FCS, 100 U/mL of penicillin, and 100 mg/mL streptomycin in 75 cm² flasks. HK cells were cultured in Epilife media, which was supplemented with HKGS. The cells were grown in a humidified incubator containing 5% CO₂ and 95% air at 37 °C until they reached 90% confluence. The following experiments were then set up for further studies concerning liposomal ATO exposure at different time intervals: cellular toxicity via MTT assay, flow cytometry analysis for cell apoptosis, and quantification of cellular arsenic uptake by inductively coupled plasma mass spectrometry (ICP-MS).