NCM460, a normal human colonic epithelial cell line, was obtained from INCELL Corporation LLC. The human colon cancer cell line HCT116 [p53 wild-type (wt)] was obtained from the Korea Cell Line Bank, and p53-null HCT116 cells were kindly provided by Dr Young-Chae Chang of the Department of Cell Biology, Catholic University of Daegu School of Medicine (Daegu, Republic of Korea). Human colon cancer DLD-1, Caco-2 and HT-29 cells were purchased from the American Type Tissue Culture Collection. The HCT116 and DLD-1 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (HyClone; GE Healthcare Life Sciences) and 1% penicillin/streptomycin (HyClone; GE Healthcare Life Sciences) at 37°C, 5% CO2. NCM460, Caco-2 and HT-29 cells were cultured in DMEM/High glucose medium (HyClone; GE Healthcare Life Sciences) supplemented in the same manner as the RPMI-1640.
Hct116
Manufactured by Korean Cell Line Bank
Sourced in United States, Cameroon
HCT116 is a human colon cancer cell line. It is a widely used in vitro model for studying various aspects of cancer biology. The core function of HCT116 is to provide a consistent and reproducible cellular system for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Ht 29, by Korean Cell Line Bank (60 mentions)
Sw480, by Korean Cell Line Bank (42 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (31 mentions)
Sw620, by Korean Cell Line Bank (23 mentions)
Dld 1, by Korean Cell Line Bank (20 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (15 mentions)
Mda mb 231, by Korean Cell Line Bank (12 mentions)
Ls174t, by Korean Cell Line Bank (10 mentions)
Mcf 7, by Korean Cell Line Bank (10 mentions)
Ccd 18co, by Korean Cell Line Bank (9 mentions)
Colo205, by Korean Cell Line Bank (8 mentions)
Caco 2, by Korean Cell Line Bank (8 mentions)
Rpmi 1640, by Welgene (8 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (8 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (8 mentions)
Dulbecco modified eagle medium (dmem), by Welgene (6 mentions)
Snu c5, by Korean Cell Line Bank (6 mentions)
Hct 15, by Korean Cell Line Bank (6 mentions)
Rpmi 1640, by Thermo Fisher Scientific (6 mentions)
Hct116, by Thermo Fisher Scientific (6 mentions)
125 protocols using hct116
1
Culturing Human Colon Cell Lines
2
Colorectal Cancer Cell Line Cultivation
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Twelve CRC cell lines (DLD1, HT29, SW620, HCT116, LoVo, KM12C, KM12SM, KM12L4, HCT116, LS174T, SW1116, and Colo205) were purchased from the Korean Cell Line Bank (https://cellbank.snu.ac.kr , Seoul, Korea). Cells were cultured in RPMI1640, MEM, DMEM, or L15 medium (Welgene, Daegu, Korea) at 37 °C in a humidified incubator with 5% CO2 according to the guidelines. All culture media contained 10% fetal bovine serum (FBS) (Gibco, Carlsbad, CA) and 1% penicillin/streptomycin (Gibco).
3
Cell Culture Protocol: Diverse Cell Lines
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4
Culturing Human Cancer Cell Lines
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Human malignant astroglioma cell line U251-MG was obtained from Dr Benveniste EN (University of Alabama at Birmingham, Birmingham, AL, USA). Human colorectal carcinoma cell line HCT116 and human malignant melanoma cell line A375 were obtained from Korean Cell Line Bank (Seoul, Korea). U251-MG were cultured in Minimum Essential Medium (MEM) (Welgene, Daegu, Korea), HCT116 in McCoy’s 5A Medium (McCoy’s) (Welgene), and A375 cells in Dulbecco’s Modified Eagle Medium (DMEM) (Welgene). Media were supplemented with 10% (v/v) fetal bovine serum (FBS) (Gibco, Logan, Utah, USA), 1x antibiotic (Welgene).
5
Colorectal Cancer Cell Lines: Cultivation and Silymarin Treatment
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Human colorectal cancer cell lines, HCT116 and SW480 were purchased from Korean Cell Line Bank (Seoul, Korea) and grown in DMEM/F-12 supplemented with 10% fatal bovine serum (FBS), 100 U/ml penicillin and 100 μg/ml streptomycin. The cells were maintained at 37°C under a humidified atmosphere of 5% CO2. Silymarin was dissolved in dimethyl sulfoxide (DMSO) and treated to cells. DMSO was used as a vehicle and the final DMSO concentration did not exceed 0.1% (v/v).
6
Cultivation and Authentication of Human Cell Lines
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The human cell lines [breast adenocarcinoma (MDA-MB-231 cells), colorectal carcinoma (HCT116 and LoVo cells), and small cell lung cancer (NCI-H889 cells)] were purchased from the Korean Cell Line Bank (Korea) and maintained in RPMI 1640 (HyClone, Korea). HEK293FT cells purchased from Invitrogen were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Hyclone). All cells were cultured in a growth medium that was supplemented with 10% heat-inactivated fetal bovine serum (FBS; HyClone), penicillin (100 U/ml), streptomycin (100 μg/ml; Welgene), and amphotericin B (0.25 μg/ml; HyClone). All cell lines were authenticated by DNA short tandem repeat profiling (ABION CRO, Korea) and used within 10 passages. All the cell lines were maintained at 37 °C in a humidified 5% CO2 incubator and were routinely screened for Mycoplasma contamination (CellSafe, Korea).
7
Colon Cancer Cell-Myofibroblast Interaction Assay
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Human colon cancer epithelial cells DLD-1 (Male, Dukes’s stage C), HCT116 (Male, Dukes’s stage D), and colonic myofibroblast CCD-18co cells (Female) were obtained from the Korean Cell Line Bank, and SW48 (Female, Dukes’s stage C) was purchased from ATCC. DLD-1 and HCT116 cells were cultured in RPMI-1640 (Gibco, Carlsbad, CA, USA) and CCD-18co and SW48 were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Gibco, Carlsbad, CA, USA) with L-glutamine (300 mg/L) supplemented with 10% fetal bovine serum (Gibco), penicillin (100 U/mL), and streptomycin (100 μM). DLD-1 cells were treated with TGFβ (R&D systems, Minneapolis, MN, USA), PDGF-D (R&D systems), imatinib (Tocris Bioscience, Bristol, UK), 2-APB (Tocris), and ML-9 (Sigma Aldrich, St. Louis, MO, USA) for 8 or 16 h. The conditioned medium was obtained from the supernatant of cells collected 8 h after PDGF-D stimulation, and treated with fresh medium in a 1:1 ratio to CCD-18co cells. To prevent mycoplasma contamination, Plasmocin™ (Invitrogen, San Diego, CA, USA, ant-mpp) was added to the medium.
8
Immunomodulatory Polymer-Based Nanoparticles
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Poloxamer 407 (pluronic F127), polyvinylpyrrolidone (PVP), dimethyl sulfoxide (DMSO), dichloromethane (DCM), 5-aminofluorescein and dialysis tube were purchased from Sigma-Aldrich. Imiquimod (R837) was purchased from Macrogen (Seoul, Korea). Dulbecco's modified eagle's medium (DMEM)-high glucose, fetal bovine serum (FBS), antibiotics were obtained from Invitrogen (Carlsbad, CA). Cell lysis buffer was used as received. Vectashield antifade mounting medium with DAPI was purchased from Vector Laboratories (Burlingame, CA). Human colorectal carcinoma cell line (HCT116) and murine macrophage cell line (RAW 264.7) were purchased from Korean Cell Line Bank (Seoul, Korea). ELISA kits were purchased from Cusabio (Wuhan, Hubei, China). Six-week-old male BALB/c mice were purchased and bred in a pathogen-free facility at the Pohang University of Science and Technology (POSTECH). All animal procedures were performed in accordance with the Guidelines for Care and Use of Laboratory Animals of POSTECH and approved by the Institutional Animal Care and Use Committee (IACUC). All chemicals were used without further purification.
9
Cell Culture and Inhibitor Treatments
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MCF-7, MDA-MB-231 and HEK 293T/17 cells (American Type Culture Collection, Manassas, VA, USA), and Colo205, Lovo, SW480, SW620, DLD-1, HCT116, HT29, LS174T and SNUC1 cells (Korean Cell Line Bank, Seoul, Republic of Korea) were cultured in Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich Corp., St. Louis, MO, USA) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (Sigma-Aldrich Corp.) and 1% (v/v) penicillin/streptomycin solution (Lonza, Basel, Switzerland) at 37 °C in a 5% CO2 incubator. In some experiments, cells were treated with specific inhibitors (Calbiochem, San Diego, CA, USA) of GSK3β (SB415286 or SB216763), p38MAPK (SB203580) or CatB (CA-074 or cathepsin inhibitor III).
10
Human Cancer Cell Culture Protocol
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