Hct15

Manufactured by American Type Culture Collection

Sourced in United States, Germany

The HCT15 is a laboratory instrument designed for the inoculation, incubation, and enumeration of cell cultures. It provides a controlled environment for the growth and maintenance of cell lines. The HCT15 allows for the precise regulation of temperature, humidity, and atmospheric composition, ensuring optimal conditions for cell cultivation.

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Lab products found in correlation

304 protocols using hct15

Culturing Colorectal Cancer Cell Lines

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CRC cell lines (HCT116, SW-620, LOVO, HCT-15 and SW480) and normal epithelial cell lines, CCD-18Co, were procured from the American Type Culture Collection (ATCC). HCT116 cells (cat. no. CCL-247) were maintained in McCoy's 5a medium (cat. no. 30-2007; ATCC); SW-620 (cat. no. CCL-227) and SW480 cells (cat. no. CCL-228) were grown in Leibovitz's L-15 medium (cat. no. 30-2008; ATCC); LOVO cells (cat. no. CCL-229) were maintained in F-12K medium (cat. no. 30-2004; ATCC); HCT-15 cells (cat. no. CCL-225) were cultured in RPMI-1640 medium (cat. no. 30-2001; ATCC); and CCD-18Co cells (cat. no. CRL-1459) were maintained in Eagle's Minimum (cat. no. 30-2003; ATCC). All cells were cultured in media supplemented with 10% FBS (cat. no. 16000044; Gibco; Thermo Fisher Scientific, Inc.) in a humidified containing 5% CO₂ at 37°C.

Ye J., Liu J., Tang T., Xin L., Bao X, & Yan Y. (2021). miR-4306 inhibits the malignant behaviors of colorectal cancer by regulating lncRNA FoxD2-AS1. Molecular Medicine Reports, 24(4), 723.

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Establishment and Characterization of CRC Cell Lines

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CRC cell lines HCT15 and Colo320 were purchased from ATCC, and HCT15/β2m cells that stably expressed the intact beta-2-microglobulin gene were established.^{33 (link)} Unless specifically mentioned, cells were cultured in complete RPMI1640 or DMEM supplemented with 10% FBS, 1% penicillin-streptomycin, 1% GlutaMAX (Gibco), 10 mM HEPES, 1 mM sodium pyruvate, and 55 µM 2-mercaptoethanol in a 5% CO2 incubator at 37°C. The T2-A24 cell line (T2 stably expressing HLA-A*24:02) was a gift from Dr. K. Kuzushima (Aichi Cancer Center Research Institute). The HLA class I genotype of HCT15 is as follows: A*24:02, A*02:01, B*08:01, B*35:01, C*04:01, and C*07:06. For gene expression and HLA genotype profiling, the data deposited in the TRON Cell Line Portal (http://celllines.tron-mainz.de) were used.^{54 (link)}

Shinkawa T., Tokita S., Nakatsugawa M., Kikuchi Y., Kanaseki T, & Torigoe T. (2021). Characterization of CD8+ T-cell responses to non-anchor-type HLA class I neoantigens with single amino-acid substitutions. Oncoimmunology, 10(1), 1870062.

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Colorectal Cancer Cell Culture Conditions

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Colorectal carcinoma cell lines HCT-116 (CCL-247), HCT-15 (CCL-225), DLD-1 (CCL-221) and hTERT RPE-1 (CRL-4000) were obtained from American Type Culture Collection and were passaged in standard conditions and recommended standard media (DLD-1 and HCT-15: RPMI1640; HCT-116: McCoy’s 5A; RPE-1: DMEM/F-12 mix) unless otherwise specified. Experimental media was prepared from powdered media (RPMI1640, Gibco 31800; McCoy’s 5A, Sigma M4892; DMEM/F-12, Thermo 12500), with the addition of 10 mM HEPES (Sigma) and 10 mM PIPES (Sigma) or 15 mM Bis-Tris (Sigma), 10% FBS (Gibco), and additional supplements to match the formula of the pre-made liquid media. 25 mM L-(+)-lactic acid (Sigma) was optionally added.

Tan Z., Chu D.Z., Chan Y.J., Lu Y.E, & Rancati G. (2018). Mammalian Cells Undergo Endoreduplication in Response to Lactic Acidosis. Scientific Reports, 8, 2890.

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Cultivation and Characterization of Human Colorectal Adenocarcinoma Cell Lines

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A total of eight human colorectal adenocarcinoma cell lines—SW-480 (CLS Cat# 300302/p716_SW-480, RRID: CVCL_0546), DLD-1 (CLS Cat# 300220/p23208_DLD-1, RRID: CVCL_0248), HCT-15 (CLS Cat# 300229/p23303_HCT-15.html, RRID: CVCL_0292), HCT-116 (CLS Cat# 300195/p19841_HCT116.html, RRID: CVCL_0291), HT-29 (NCI-DTP Cat# HT-29, RRID: CVCL_0320), LS174T (CLS Cat# 300392/p720_LS-174T, RRID: CVCL_1384), SW-620 SW620 (NCI-DTP Cat# SW-620, RRID: CVCL_0547) and LoVo (NCI-DTP Cat# LOVO, RRID: CVCL_0399)—were obtained from the American Type Culture Collection (ATCC, Rockville, Maryland). The cell lines were cultivated at 37 °C in high-glucose Dulbecco’s modified Eagle’s medium (DMEM) containing 10% heat-inactivated fetal bovine serum (FBS), 50 U/mL penicillin, and 50 µg/mL streptomycin (Sigma-Aldrich Chemical Co., USA) in an atmosphere of 5% CO₂ and 95% humidified air. Subculture was performed when 90% confluence was reached. Human cell line identification by short tandem repeat profile testing, according to the American National Standards Institute, has shown an appropriate match for HCT-116 and DLD-1 cell lines.

Alburquerque-González B., Bernabé-García M., Montoro-García S., Bernabé-García Á., Rodrigues P.C., Ruiz Sanz J., López-Calderón F.F., Luque I., Nicolas F.J., Cayuela M.L., Salo T., Pérez-Sánchez H, & Conesa-Zamora P. (2020). New role of the antidepressant imipramine as a Fascin1 inhibitor in colorectal cancer cells. Experimental & Molecular Medicine, 52(2), 281-292.

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Novel cytotoxic analogs of KTH-13 compound

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4-Methyl-2,6-bis(1-phenylethyl)phenol [KTH-13-Me] and its structural analogs, including KTH-13-benzyl-glycol, KTH-13-monophenylethyl-glycol, KTH-13-monobenzyl-glycol, KTH-13-monophenylethyl, KTH-13-monobenzyl-FA, KTH-13-monophenyle thyl-FA, and KTH-13-monophenylethyl-FA (Fig. 1) were supplied by Prof Lee, Yunmi (Kwangwoon University, Seoul, Korea). Hoechst stain solution, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), sodium dodecyl sulfate (SDS), staurosporine, and dimethyl sulfoxide (DMSO) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Fetal bovine serum (FBS), penicillin, and streptomycin were obtained from GE Healthcare Hyclone (Grand Island, NY, USA). C6 glioma, MDA-MB 231, HCT-15, and LoVo cells were purchased from ATCC (Rockville, MD, USA). Antibodies to phospho-, cleaved- or total-protein forms of Src, STAT3, caspase-3, caspase-9, Bcl-2, and β-actin were obtained from Cell Signaling Technology (Beverly, MA, USA).

Sung N.Y., Kim S.C., Kim Y.H., Kim G., Lee Y., Sung G.H., Kim J.H., Yang W.S., Kim M.S., Baek K.S., Kim J.H, & Cho J.Y. (2016). Anti-Proliferative and Pro-Apoptotic Activities of 4-Methyl-2,6-bis(1-phenylethyl)phenol in Cancer Cells. Biomolecules & Therapeutics, 24(4), 402-409.

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Culturing Colon and Breast Cancer Cells

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Colon adenocarcinoma (HCT-15) and breast adenocarcinoma (MCF-7) were obtained from ATCC. Medium contained 10% fetal bovine serum, penicillin 100 IU/mL, and streptomycin 100 g/mL, RPMI, at 37°C, 5% CO₂, and water saturated humidity condition. Every 1 or 2 days, for a fluid passage, the logarithm growth period of cell activity of more than 95% cells was used in this study.

Bolaños-Carrillo M.A., Ventura-Gallegos J.L., Saldivar-Jiménez A.D., Zentella-Dehesa A, & Martínez-Vázquez M. (2015). Effect of Sterols Isolated from Myrtillocactus geometrizans on Growth Inhibition of Colon and Breast Cancer Cells. Evidence-based Complementary and Alternative Medicine : eCAM, 2015, 589350.

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Authenticated Cell Lines for Protein Interaction Studies

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The following authenticated cell lines were purchased from ATCC: MDCK.2 (CRL-2936, RRID:CVCL_B034), HEK293 (CRL-1573, RRID:CVCL_0045), and human colon cancer lines including HT29 (HTB-38, RRID:CVCL_0320), HCT116 (CCL-247, RRID:CVCL_0291), HCT15 (CCL-225, RRID:CVCL_0292), SW480 (CCL-228, RRID:CVCL_0546), Caco2 (HTB-37, RRID:CVCL_0025), SW620 (CCL-227, RRID:CVCL_0547), Ls174T (CL-188, RRID:CVCL_1384), and T84 (CCL-248, RRID:CVCL_0555). Cells were cultured following the manufacturer's guidelines, and passaged up to five times after each thawing. All cell lines were routinely tested for Mycoplasma contamination using Universal Mycoplasma Detection Kit (ATCC, 30–1012K). HEK293 cells were transfected with equimolar amounts of control empty plasmid or plasmid encoding FILIP1L-HA and/or Flag-PFDN1 using lipofectamine 3000 solution (Thermo Fisher Scientific) following the manufacturer's protocols. For MDCK.2 cells, using the 4D-Nucleofector system (Lonza; SE solution; program CM 113), homogenous expression in over 90% of cells was routinely achieved. At 24 to 48 hours following transfection, transfected cells were subjected to downstream assays such as immunoprecipitation, immunoblot, and immunofluorescence staining.

Kwon M., Rubio G., Nolan N., Auteri P., Volmar J.A., Adem A., Javidian P., Zhou Z., Verzi M.P., Pine S.R, & Libutti S.K. (2021). FILIP1L Loss Is a Driver of Aggressive Mucinous Colorectal Adenocarcinoma and Mediates Cytokinesis Defects through PFDN1. Cancer Research, 81(21), 5523-5539.

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Colon Cancer Cell Lines and Non-cancerous Tissue RNA

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Four human colon cancer cell lines, HCT-15, HT-29, SK-CO-1 and WiDr (ATCC, Manassas, VA) and total RNA samples isolated from two independent sources of non-cancerous colon tissues (Origene, Rockville, MD) were used in this work.

Choo K.B., Soon Y.L., Nguyen P.N., Hiew M.S, & Huang C.J. (2014). MicroRNA-5p and -3p co-expression and cross-targeting in colon cancer cells. Journal of Biomedical Science, 21(1), 95.

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CRC Cell Culture and Tissue Procurement

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The CRC cells used in this study including HCT116, HT-29, LoVo, SW480, Caco2, HCT-15, RKO and HUVECs were purchased from ATCC. Cells were maintained in RPMI 1640 (Gibco-BRL, Germany) supplemented with 10% fetal bovine serum (FBS, Biological Industries BI, Israel). Cells were cultured in a moist environment containing 5% CO₂ at 37 °C. CRC tissues and paired surgical margin tissues were obtained after surgical procedures conducted at Chongqing University Cancer Hospital, Chongqing, China.

Xiang Q., Zhou D., Xiang X., Le X., Deng C., Sun R., Li C., Pang H., He J., Zheng Z., Tang J., Peng W., Peng X., He X., Wu F., Qiu J., Xu Y, & Xiang T. (2023). Neuroglobin plays as tumor suppressor by disrupting the stability of GPR35 in colorectal cancer. Clinical Epigenetics, 15, 57.

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Cytotoxicity Evaluation of Human Colon Cancer Cell Lines

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For cytotoxicity tests, three different human colon cancer cell lines were used: HCT15 and HCT116 were purchased from the ATCC (American Type Culture Collection). HCT116oxR was established by one of the authors (U. J.) at the Institute of Cancer Research, Department of Medicine I, Medical University Vienna, Austria, as described in ref. 46 (link). Every 3–4 passages, the cells were treated with 10 μM oxaliplatin for 3 days to ensure maintenance of the resistance. All cell culture media and reagents were purchased from Sigma-Aldrich Austria and all cell culture materials such as dishes, plates and flasks from StarLab Germany unless indicated otherwise. Cells were grown as adherent monolayer cultures in 75 cm² culture flasks in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum (Invitrogen) and 4 mM l-glutamine without antibiotics at 37 °C under a humid atmosphere containing 5% CO₂ and 95% air. The murine colon cancer cell line CT26 and murine leukemia L1210 (both from ATCC) were grown in DMEM/F12 or RMPI 1640 medium, respectively, supplemented with 10% heat-inactivated fetal bovine serum.

Göschl S., Schreiber-Brynzak E., Pichler V., Cseh K., Heffeter P., Jungwirth U., Jakupec M.A., Berger W, & Keppler B.K. (2017). Comparative studies of oxaliplatin-based platinum(iv) complexes in different in vitro and in vivo tumor models. Metallomics : integrated biometal science, 9(3), 309-322.

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