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Hct116

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HCT116 is a human colorectal carcinoma cell line. It is a commonly used in vitro model for cancer research.

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Lab products found in correlation

Hct116, by American Type Culture Collection (19 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (13 mentions) Dulbecco modified eagle medium (dmem), by Corning (12 mentions) Fetal bovine serum (fbs), by Corning (10 mentions) Sw480, by Corning (8 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (8 mentions) Sw620, by Corning (7 mentions) Rpmi 1640, by Corning (6 mentions) Dld 1, by American Type Culture Collection (5 mentions) Mccoy s 5a, by Corning (5 mentions) Ht 29, by Corning (5 mentions) Rpmi 1640 medium, by Corning (5 mentions) Penicillin streptomycin, by Corning (4 mentions) Hek293t, by Corning (4 mentions) Mycoalert mycoplasma detection kit, by Lonza (3 mentions) Sw480, by American Type Culture Collection (3 mentions) Dmem f12, by Corning (3 mentions) Mccoy s 5a medium, by Corning (3 mentions) Mcf 7, by Corning (3 mentions) Mda mb 468, by Corning (3 mentions)

40 protocols using hct116

1

Culturing a CRC cell line panel

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A panel of CRC cell lines was purchased (ATCC) and grown in the vendor-recommended media: DLD-1 (RPMI-1640, Cellgro); HT29 (MacCoy’s 5a modified with 2% NaHCO3, Cellgro); HCT116 (MacCoy’s 5a, Cellgro); Colo201 (RPMI-1640 modified with 1% sodium pyruvate, Cellgro); SW480 and SW620 (Dulbecco’s modified essential medium, Cellgro); CCD-18Co (Eagle’s minimum essential medium, Cellgro); CCD-112CoN (Eagle’s minimum essential medium, Cellgro); CCD-33Co (Eagle’s minimum essential medium, Cellgro). All media were supplemented with 10% FBS and penicillin-streptomycin (Cellgro). All cell lines were tested and determined to be free of mycoplasma contamination (MycoAlert Mycoplasma Detection Kit, Lonza, LT07-418).
Park J., Park J.S., Huang C.H., Jo A., Cook K., Wang R., Lin H.Y., Deun J.V., Li H., Min J., Wang L., Yoon G., Carter B.S., Balaj L., Choi G.S., Castro C.M., Weissleder R, & Lee H. (2021). A high-throughput magneto-electrochemical array for the integrated isolation and profiling of extracellular vesicles from plasma. Nature biomedical engineering, 5(7), 678-689.
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2

Colon Cancer Cell Lines and Culture Conditions

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Colon cancer cell lines LOVO, HCT116, Caco-2, HT29, SW480, and SW1116 were obtained from American Type Culture Collection (ATCC) and subcultured and preserved by Shanghai Institute of Digestive Surgery; normal human colon mucosal epithelium cell NCM460 was kindly donated by Jingqing Zeng (Ruijin Hospital, Shanghai Jiao Tong University School of Medicine; the donator bought NCM460 cell line from INCELL Corporation, LLC, San Antonio, TX, USA). LOVO was cultured in F-12K Nutrient Mixture Medium (Corning Cellgro®, MA, USA), HCT116 and HT29 in McCoy's 5A (Corning Cellgro®), SW480 and SW1116 in Leibovitz' L-15 (Corning Cellgro®), and NCM460 in DMEM (Corning Cellgro®). All culture medium above was supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Carlsbad, CA, USA). Caco-2 was cultured in Minimum Essential Medium (Corning Cellgro®) with 20% FBS. Cells were placed in the incubator (Heracelles, Germany) at 37°C, in a humidified atmosphere with 5% CO2.
Huang A., Zhao H., Quan Y., Jin R., Feng B, & Zheng M. (2014). E2A Predicts Prognosis of Colorectal Cancer Patients and Regulates Cancer Cell Growth by Targeting miR-320a. PLoS ONE, 9(1), e85201.
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3

Culturing and Maintaining Human Colorectal Cancer Cell Lines

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HCT-116 (CCL-247), LoVo (CCL-229), DLD-1 (CCL-221), HT-29 (HTB-38) and CoN (CCD-841) human CRC cells were obtained from the ATCC (Manassas, VA, USA). HCT-116 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Corning Cellgro, Corning, NY, USA) for the invasion assays or RPMI-1640 medium (Lonza, Westburg, The Netherlands) for all other assays. The LoVo, DLD-1, and HT-29 cell lines were grown in RPMI-1640 for all experiments. Media were supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA or Greiner Bio-One, Vilvoorde, Belgium), 100 U/mL penicillin, and 100 µg/mL streptomycin (Gibco, Waltham, MA). Cells were grown at 37 °C and 5% CO2 in a humidified atmosphere. The CoN cells were cultured in Eagle’s Minimum Essential Medium (Lonza, Westburg, The Netherlands) supplemented with 10% FBS (South America origin, Greiner Bio-One, Vilvoorde, Belgium). HCT-116 p53-/- cells were obtained from Denis L.J. LaFontaine, Université Libre de Bruxelles, Brussels, Belgium [42 (link)].
Mathieu V., Laguera B., Masi M., Dulanto S.A., Bingham T.W., Hernandez L.W., Sarlah D., Evidente A., Lafontaine D.L., Kornienko A, & Lane M.A. (2022). Amaryllidaceae Alkaloids Decrease the Proliferation, Invasion, and Secretion of Clinically Relevant Cytokines by Cultured Human Colon Cancer Cells. Biomolecules, 12(9), 1267.
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4

Culturing a CRC cell line panel

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A panel of CRC cell lines was purchased (ATCC) and grown in the vendor-recommended media: DLD-1 (RPMI-1640, Cellgro); HT29 (MacCoy’s 5a modified with 2% NaHCO3, Cellgro); HCT116 (MacCoy’s 5a, Cellgro); Colo201 (RPMI-1640 modified with 1% sodium pyruvate, Cellgro); SW480 and SW620 (Dulbecco’s modified essential medium, Cellgro); CCD-18Co (Eagle’s minimum essential medium, Cellgro); CCD-112CoN (Eagle’s minimum essential medium, Cellgro); CCD-33Co (Eagle’s minimum essential medium, Cellgro). All media were supplemented with 10% FBS and penicillin-streptomycin (Cellgro). All cell lines were tested and determined to be free of mycoplasma contamination (MycoAlert Mycoplasma Detection Kit, Lonza, LT07-418).
Park J., Park J.S., Huang C.H., Jo A., Cook K., Wang R., Lin H.Y., Deun J.V., Li H., Min J., Wang L., Yoon G., Carter B.S., Balaj L., Choi G.S., Castro C.M., Weissleder R, & Lee H. (2021). A high-throughput magneto-electrochemical array for the integrated isolation and profiling of extracellular vesicles from plasma. Nature biomedical engineering, 5(7), 678-689.
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5

Cell Culture Protocols for Biomedical Research

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The human monocyte THP-1 (ATCC TIB-202) cell line; the human colon epithelial cell lines HT-29, HCT-116 and SW 620; and the mouse macrophage RAW 264.7 cell line were purchased from the American Type Culture Collection (Manassas, VA). HT-29, HCT-116, SW 620 and RAW 264.7 cells were cultured in complete growth media (CGM) using Dulbecco’s Modified Eagle’s Medium (DMEM) with 4.5 g/L glucose, L-glutamine and sodium pyruvate (Cellgro, Corning, NY) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Atlanta Biologics, Lawrenceville, GA) and 1% penicillin-streptomycin (Cellgro, Corning, NY). THP-1 cells were grown in Roswell Park Memorial Institute (RPMI) medium with L-glutamine (Cellgro, Corning, NY) supplemented with 10% heat-inactivated FBS and 1% penicillin-streptomycin. All cells were maintained in an incubator set at 37°C and 5% CO2 and allowed to grow to 80–90% confluency before being sub-cultured or used in experiments. Purity of DMA was confirmed by gas chromatography-mass spectroscopy. All other reagents were purchased from Sigma-Aldrich (St. Louis, MO) or VWR (Bridgeport, NJ).
Koya J.B., Shen T., Lu G., Gauthier A., Mantell L., Ashby CR J.r, & Reznik S.E. (2022). FDA-Approved Excipient N, N-Dimethylacetamide Attenuates Inflammatory Bowel Disease in In Vitro and In Vivo Models. Fortune journal of health sciences, 5, 499-509.
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6

Authenticated Cell Line Comparison for Research

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HCT116 and HCT116 ARID1A-/- cells were purchased from Horizon Discovery with cell line authentication information provided (STR Profile: Amelogenin: X, Y; CSF1PO: 7, 10; D13S317: 10, 12; D16S539: 11, 13; D5S818: 10, 11; D7S820: 11, 12; THO1: 8, 9; TPOX: 8, 9; vWA: 17,22; ATCC). Both lines were used within six months of receipt. HCT116 ARID1A-/- cells were generated by knock-in of a premature stop codon at Q456 (Horizon Discovery). HCT116 and HCT116 ARID1A-/- cells were grown in RPMI1640 media (Corning CellGro) supplemented with 10% FBS (Omega Scientific, Inc.) and 1% Penicillin/Streptomycin (Life Technologies). TOV21G cells were purchased from ATCC with cell line authentication information provided (STR Profile: Amelogenin: X; CSF1PO: 13, 15; D13S317: 11, 12; D16S539: 10, 12; D5S818: 12, 13; D7S820: 12; THO1: 7, 9.3; TPOX: 8, 11; vWA: 17; ATCC). Cells were used within six months of receipt. TOV21G cells were grown in 1:1 mixture of Medium 199 (Life Technologies) containing 2.2 g/L sodium bicarbonate and MCDB 105 (Life Technologies) containing 1.5 g/L sodium bicarbonate, supplemented with 15% FBS and 1% Penicillin/Streptomycin. All cells were grown at 37˚C with 5% CO2. All cell lines were negative for mycoplasma when tested by MycoAlert Mycoplasma Detection Kit (Lonza, Basel, Switzerland)
Kelso T.W., Porter D.K., Amaral M.L., Shokhirev M.N., Benner C, & Hargreaves D.C. (2017). Chromatin accessibility underlies synthetic lethality of SWI/SNF subunits in ARID1A-mutant cancers. eLife, 6, e30506.
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7

Colon and Stomach Cell Line Treatments

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All cell lines were maintained in a humidified atmosphere with 5% CO2. Our study included five colon cell lines (HT29, SW480, HCT116, LoVo and RKO) and one stomach cell line (AGS). HT29, SW480, HCT116 and LoVo cells were cultured in McCoys 5A media (Corning), RKO and AGS were cultured in RPMI 1640 media (Corning) supplemented with 10% FBS (Gibco). All cell lines were purchased from the ATCC and authenticated and tested for Mycoplasma by IDEXX on 6/20/2019. All cells used in experiments were passaged fewer than 15 times with most being passaged fewer than 10 times. For H2O2 treatments, 30% H2O2 (Sigma) was diluted in PBS immediately prior to treatment at 250 μM for 1H at 37°C. For EGF treatments, cells were starved in media lacking serum for 48H prior to treatment. Cells were then treated with 100 ng/ml recombinant EGF (R&D Systems: 236-EG) for 48H. GSK-LSD1 (Sigma, SML1072), GSK690693 (Sigma, SML0428) and corin (generously provided by Dr. Philip Cole and Dr. Jay Kalin) were solubilized in DMSO (Sigma) prior to treatment. Treatment dosages and durations are defined in the figure legends.
Miller S.A., Policastro R.A., Savant S.S., Sriramkumar S., Ding N., Lu X., Mohammad H.P., Cao S., Kalin J.H., Cole P.A., Zentner G.E, & O’Hagan H.M. (2019). Lysine-specific demethylase 1 mediates AKT activity and promotes epithelial-mesenchymal transition in PIK3CA mutant colorectal cancer. Molecular cancer research : MCR, 18(2), 264-277.
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8

Culture of Colorectal Cancer Cell Lines

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Colorectal cancer cell lines HCT116 and HT29 were purchased from KeyGEN BioTECH. CO., LTD (Nanjing, China). HCT116 cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Corning) supplemented with 10% fetal bovine serum (FBS, CellMax) and 1% PS (penicillin-streptomycin, Gibco), and HT29 cells were cultured in RPMI 1640 medium (KeyGEN) supplemented with 10% FBS (Biological Industries) and 1% PS, and maintained in a humidified atmosphere containing 5% CO2 at 37 ºC.
Liang X., Lan C., Zhou J., Fu W., Long X., An Y., Jiao G., Wang K., Li Y., Xu J., Huang Q., Xu B, & Xiao J. (2017). Development of a new analog of SGK1 inhibitor and its evaluation as a therapeutic molecule of colorectal cancer. Journal of Cancer, 8(12), 2256-2262.
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9

Cell Culture Conditions and Synchronization

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HeLa, HCT116, hTERT-RPE1, A549 and U2OS cells (American Type Culture Collection, Manassas, VA) were cultured in a humidified incubator at 37 °C in the presence of 5% CO2. For hTERT-RPE1 and U2OS cells, stable cell lines expressing H2B–GFP were derived using described methods45 . HeLa, HCT116 and A549 cells were grown in DMEM (10–013-CV, Corning) with 10% FBS. hTERT-RPE1 H2B-GFP cells were grown in DMEM/F12 (10–090-CV, Corning) with 10% FBS and 0.01 mg/ml hygromycin B. U2OS H2B-GFP cells were grown in McCoy’s 5A (10–050-CV, Corning) with 10% FBS.
Unless otherwise noted, HeLa and HCT116 cells were synchronized by treating with 2 mM thymidine for 18 hours, releasing for 8 hours and retreating for 18 hours. In all cellular experiments involving chemicals dissolved in DMSO, the final DMSO concentration was kept below 0.5%.
Richeson K.V., Bodrug T., Sackton K.L., Yamaguchi M., Paulo J.A., Gygi S.P., Schulman B.A., Brown N.G, & King R.W. (2020). Paradoxical mitotic exit induced by a small molecule inhibitor of APC/CCdc20. Nature chemical biology, 16(5), 546-555.
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10

Colorectal and Kidney Cell Lines

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The human colorectal cancer cell line HCT116, SW480, DLD1 and LOVO, as well as the human embryonic kidney cell lines HEK-293T were purchased from ATCC. All cells were grown in RPMI (Corning, 10–040-CV) supplemented with 10% fetal bovine serum (Bioexpress, S1200–500), and penicillin and streptomycin (Lonza, 17–602E). The doxycycline-inducible pINDUCER HCT116, SW480, DLD1, and LOVO cell lines were maintained in the same condition with tetracycline-free fetal bovine serum (Corning, #35–075-CV) and periodically selected by supplemental 500 μg/mL G418 (Sigma, #A1720).
Chen M.S., Lo Y.H., Chen X., Williams C.S., Donnelly J.M., Criss ZK I.I., Patel S., Butkus J.M., Dubrulle J., Finegold M.J, & Shroyer N.F. (2019). Growth Factor–Independent 1 Is a Tumor Suppressor Gene in Colorectal Cancer. Molecular cancer research : MCR, 17(3), 697-708.
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