The following source of the cell lines were COLO 201, HT29, HCT 116, SW837, SW620, and SW1463 (ATCC); LIM2551, LIM1215, and LIM2099 (Ludwig Institute for Cancer Research, Melbourne, Australia), GEO (29 (link)), DIFI (30 (link)), and V9P (31 (link)). All cell lines were maintained in DMEM/F12 supplemented with 5% FBS (v/v; all from Thermo Fisher Scientific) at 37°C with 5% CO2. Cell lines were authenticated by short tandem repeat profiling using the GenePrint 10 system (Promega), and all were found to be exact matches to published profiles. All cell lines were frozen down as large batches of master stocks within five passages of their purchase from these commercial vendors and confirmed to be Mycoplasma negative at the point of freezing. Experiments were then performed using these master stocks for up to 20 passages. Mycoplasma testing was performed every 3 to 6 months as part of routine monitoring in our laboratory.
Sw837
Manufactured by American Type Culture Collection
Sourced in United States
The SW837 is a laboratory centrifuge designed for general-purpose applications. It features a compact and durable construction, quiet operation, and a wide range of speed and acceleration options. The SW837 is suitable for a variety of sample processing tasks in life science research and clinical laboratories.
Automatically generated - may contain errors
Lab products found in correlation
Hct116, by American Type Culture Collection (42 mentions)
Sw480, by American Type Culture Collection (34 mentions)
Sw620, by American Type Culture Collection (30 mentions)
Sw1463, by American Type Culture Collection (20 mentions)
Dld 1, by American Type Culture Collection (18 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (17 mentions)
Caco 2, by American Type Culture Collection (15 mentions)
Hct15, by American Type Culture Collection (13 mentions)
Sw1116, by American Type Culture Collection (13 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (12 mentions)
Ls174t, by American Type Culture Collection (11 mentions)
Sk co 1, by American Type Culture Collection (10 mentions)
Sw837, by Thermo Fisher Scientific (10 mentions)
Sw948, by American Type Culture Collection (9 mentions)
Sw480, by Thermo Fisher Scientific (9 mentions)
Hct116, by Thermo Fisher Scientific (9 mentions)
Colo205, by American Type Culture Collection (8 mentions)
Sw620, by Thermo Fisher Scientific (8 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (7 mentions)
Ls123, by American Type Culture Collection (7 mentions)
72 protocols using sw837
1
Characterization of Colorectal Cancer Cell Lines
2
Evaluating BET Inhibitor Cytotoxicity in CRC
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All CRC cell lines (HT29, DLD1, Colo205, H716, SW837, H508 and MC38) used in this study were purchased from ATCC and kept in our laboratory. The CellTiter-Glo method was used in this experiment. CRC cells were plated in cell culture plates and cultured in a cell culture incubator at 37 °C, 5% CO2 or 100% air and 95% humidity. Compounds were added the next day and incubated for 5 d, and cell viability was detected with the CellTiter-Glo kit. The data were analyzed using GraphPad Prism software, and a four-parameter equation was used to fit a concentration-response curve, from which the IC50 of the compound concentration corresponding to 50% cell viability on the curve was calculated. Cell viability (%) = (Lumitest compound-Lumiblank control)/(Lumisolvent control-Lumiblank control) × 100%. Compound information: BET inhibitor JAB-8263 (Jacobio Pharmaceuticals, Beijing, China), purity: 99.10%, storage condition: 4 °C.
3
Colorectal Cancer Cell Line Culture
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The rectal cancer cell lines SW837 and SW1463">SW1463 and the colon cancer cell lines HCT116 and LS513 were obtained from the ATCC. Rectal and colon cancer cell lines were cultured in Leibovitz's 15 (Welgene) at 37°C in an atmosphere containing 1% CO2 or in RPMI 1640 (Lonza) at 37°C in an atmosphere containing 5% CO2 containing 10% FBS (Corning) and gentamycin (50 μg/mL; Lonza), respectively. 5‐Fluorouracil (5‐FU), TGFBR inhibitor (SB525334 ), metformin and phenformin were obtained from Sigma‐Aldrich. The signal transducer and activator of transcription 3 (STAT3) inhibitor STATTIC was purchased from Calbiochem. For assessing apoptosis, apoptotic protein assay (ARY009) was purchased and was performed according to the instructions provided using the R&D system.
4
Characterization of CRC Cell Lines
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Fifteen CRC cell lines were studied: the near diploid cell lines DLD-1, HCT116,
HCT116p53-/-, SW48, and LoVo (all from ECACC except HCT116p53-/- which was a gift
from Dr G Smith, University of Dundee, UK [18 (link)]) and the aneuploid cell lines SW480, SW837, HT29, T84,
Colo 201, Colo 320DM, LS411N, SK-CO-1, NCI H508 and NCI H716 (all from ATCC) apart
from Colo 320DM, T84, and SW837 (all from ECACC).
The cell lines were cultured in Dulbecco’s modified Eagle’s medium
(DMEM) (Gibco®, Cat. no. 31885) supplemented with 10% foetal bovine
serum (FBS; PAA, Cat. no. A15-101) and 1% penicillin-streptomycin
(Gibco®, Cat. no.15140-122). The cell lines were grown in a
humidified incubator at 37°C containing 5% CO2. All the cell lines
were tested for mycoplasma using the Venor™GeM Mycoplasma Detection
Kit (Sigma-Aldrich, Cat. no. MP0025). When the cell lines reached 70–80%
confluence, they were trypsinized using 0.05% trypsin-EDTA (1X) with phenol red
(Gibco®, Cat. no. 25300).
HCT116p53-/-, SW48, and LoVo (all from ECACC except HCT116p53-/- which was a gift
from Dr G Smith, University of Dundee, UK [18 (link)]) and the aneuploid cell lines SW480, SW837, HT29, T84,
Colo 201, Colo 320DM, LS411N, SK-CO-1, NCI H508 and NCI H716 (all from ATCC) apart
from Colo 320DM, T84, and SW837 (all from ECACC).
The cell lines were cultured in Dulbecco’s modified Eagle’s medium
(DMEM) (Gibco®, Cat. no. 31885) supplemented with 10% foetal bovine
serum (FBS; PAA, Cat. no. A15-101) and 1% penicillin-streptomycin
(Gibco®, Cat. no.15140-122). The cell lines were grown in a
humidified incubator at 37°C containing 5% CO2. All the cell lines
were tested for mycoplasma using the Venor™GeM Mycoplasma Detection
Kit (Sigma-Aldrich, Cat. no. MP0025). When the cell lines reached 70–80%
confluence, they were trypsinized using 0.05% trypsin-EDTA (1X) with phenol red
(Gibco®, Cat. no. 25300).
5
Cell Culture Protocol for Cystine Restriction
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SW620, SW837, SW480, DLD1, and HEK293T cells were purchased from the ATCC. Wild-type and CBS KO MEFs were generated from day 13.5 embryos isolated from wild-type and CBS KO mice (C57B6, #002461, The Jackson Laboratory). DLD1 cells were cultured in RPMI-1640 medium (Shanghai Basal Media Technologies Company, L210KJ). HEK293T cells and MEFs were cultured in DMEM (Shanghai Basal Media Technologies Company, L110KJ). SW480, SW620, and SW837 cells were cultured in L-15 medium (Shanghai Basal Media Technologies Company, L620KJ). For cystine restriction experiments, cells were grown in L-cystine-free RPMI 1640 medium or in medium supplemented with 65 mg/L L-cystine. All cells were grown in a medium containing 10% fetal bovine serum (FBS; A24G00J, Gemini) supplemented with 100 units/mL penicillin and streptomycin (PSL01, Caisson). All cells were incubated at 37 °C in a humidified atmosphere containing 5% CO2. All cell lines were identified using short tandem repeat (STR) profiling.
6
Culturing Human Colon and Colorectal Cancer Cells
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Human colon (HCT116 and HT-29) and colorectal (SW-837) adenocarcinoma cell lines were purchased from ATCC. All cancer cells were cultured in RPMI-1640 medium with ultraglutamine (Lonza, VWR, Carnaxide, Portugal), and supplemented with 10% fetal bovine serum (FBS; Gibco, Alfagene, Carcavelos, Portugal). Cells were maintained at 37 °C in a humidified atmosphere of 5% CO2.
7
Culturing Colon and Rectal Cancer Cells
8
Characterization of CRC Cell Lines
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The human colorectal tumor cell lines HCT116, DLD1, SW480, SW620, SW837, and HT29 were obtained from ATCC. HCT116 cells line was used as in vitro model due to wildtype Tp53 status compared to other CRC cell lines (Supplementary Table 1 ). HCT116 and HCT116-derived monoclonal DAPK1 ko clones 7/6, 10/8 and 21/9 were maintained in RPMI 1640 medium supplemented with 10% (v/v) fetal bovine serum (FBS), 1% penicillin (100 U/ml) and streptomycin (100 µg/ml) (all from PAN Biotech, Aidenbach, Germany) and cultured in a humidified atmosphere of 5% CO2 at 37 °C. For ERK1/2 inactivation, cells were treated with 10 and 20 µM FR180204 (Selleckchem, Munich, Germany) for 48 h. All cell lines were genotyped using Multiplex Cell Authentication by Multiplexion (Heidelberg, Germany) as described recently15 (link). Mycoplasma-free status has been verified for all cell lines.
9
Comprehensive Cell Line Authentication Protocol
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All cell lines, including HPAC, MIA Paca-2, Capan-1, NCI-H2122, NCI-H2030, SW837, SW1463 and 293 T cells, were purchased from ATCC and authenticated on their own cell lines. Pa01c, Pa02c, Pa03c, Pa14c, and Pa16c were kind gifts from Dr. Channing Der at UNC-CH and whole-exome sequenced72 (link). All lines were used for <6 months after receipt or resuscitation from cryopreservation. All cell lines were cultured in DMEM supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. Mycoplasma testing was performed semiannually using a MycoSEQ Detection kit (Applied Biosystems).
10
Regulation of Rectal Cancer by miR-195
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Human rectal mucosa epithelial cell line (PriCells) and 2 types of human rectal cancer cell lines (SW837 and SW1463; ATCC) were cultured with Leibovitz's 15 (L-15) culture medium (ATCC), which was supplemented with 10% fetal bovine serum (FBS; ATCC) in 5% CO2 in an incubator at 37°C. The cells in logarithmic phase were harvested for subsequent experiments.
miR-195 mimic (100 pmol) (sense, 5′-UAGCAGCACAGAAAUAUUGGC-3′ and antisense, 5′-CAAUAUUUCUGUGCUGCUAUU-3′) and mimic control (sense, 5′-UUCUCCGAACGUGUCACGUTT-3′ and antisense, 5′-ACGUGACACGUUCGGAGAATT-3′) were obtained from Shanghai GenePharma Co., Ltd. Full-length insulin-like growth factor 1 (IGF1 sense, 5′-GAATTCATGGGAAAAATCAGCAGTC-3′ and antisense, 5′-GATATCGCATGTCATTCTTCACTCTTT-3′) were cloned into a pcDNA3.1 vector (Takara Biotechnology Co., Ltd.), and an empty pcDNA3.1 was set as negative control (NC). Lipofectamine 3000 (Invitrogen; Thermo Fisher Scientific, Inc.) was used in cell transfection. After being transfected with miR-195mimic, mimic control, IGF1 or NC vectors for 48 h, the cells were transferred to complete medium containing puromycin.
miR-195 mimic (100 pmol) (sense, 5′-UAGCAGCACAGAAAUAUUGGC-3′ and antisense, 5′-CAAUAUUUCUGUGCUGCUAUU-3′) and mimic control (sense, 5′-UUCUCCGAACGUGUCACGUTT-3′ and antisense, 5′-ACGUGACACGUUCGGAGAATT-3′) were obtained from Shanghai GenePharma Co., Ltd. Full-length insulin-like growth factor 1 (IGF1 sense, 5′-GAATTCATGGGAAAAATCAGCAGTC-3′ and antisense, 5′-GATATCGCATGTCATTCTTCACTCTTT-3′) were cloned into a pcDNA3.1 vector (Takara Biotechnology Co., Ltd.), and an empty pcDNA3.1 was set as negative control (NC). Lipofectamine 3000 (Invitrogen; Thermo Fisher Scientific, Inc.) was used in cell transfection. After being transfected with miR-195mimic, mimic control, IGF1 or NC vectors for 48 h, the cells were transferred to complete medium containing puromycin.
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