Hct 8

Manufactured by American Type Culture Collection

Sourced in United States

The HCT-8 is a laboratory equipment designed for cell culture applications. It is a small-scale bioreactor system that provides a controlled environment for the growth and maintenance of various cell lines. The HCT-8 is capable of regulating temperature, pH, and dissolved oxygen levels to support the optimal conditions for cell proliferation and differentiation.

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Lab products found in correlation

Close spelling variants of this product

HCT-8 cells (26 citations) HCT-8 (CCL-244) (6 citations) HCT-8 human ileocecal adenocarcinoma cells (2 citations)

73 protocols using hct 8

Cultivation and Titration of SARS-CoV-2 and Common Coronaviruses

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Human lung fibroblasts MRC5 (ATCC CCL-171), the human colorectal carcinoma HCT-8 (ATCC CCL-244), the human lung adenocarcinoma Calu-3 (ATCC HTB-55), and the African green monkey kidney Vero E6 (ATCC CRL-1586) cell lines were purchased from the American Type Culture Collection (ATCC), and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Euroclone) supplemented with 10% fetal bovine serum (FBS, Euroclone), 2 mM glutamine, 1 mM sodium pyruvate, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate (P/S, both from Euroclone).
hCoV-229E (ATCC VR-740) and hCoV-OC43 (ATCC VR-1558) were purchased from ATCC, and propagated and titrated in MRC5 and HCT-8 cells, respectively. SARS-CoV-2 (2019-nCoV/Italy INMI1) was obtained from EVAg, and propagated and titrated in Vero E6 cells. SARS-CoV-2/01/human/2020/SWE was isolated on Vero E6 cells from a nasopharyngeal sample, cultivated and titrated as previously described [18 (link)]. All work with SARS-CoV-2 was performed in Biosafety laboratory level 3 (BSL3) facilities either at the University of Padua, Italy, or at the Public Health Agency of Sweden, Sweden.

Calistri A., Luganini A., Mognetti B., Elder E., Sibille G., Conciatori V., Del Vecchio C., Sainas S., Boschi D., Montserrat N., Mirazimi A., Lolli M.L., Gribaudo G, & Parolin C. (2021). The New Generation hDHODH Inhibitor MEDS433 Hinders the In Vitro Replication of SARS-CoV-2 and Other Human Coronaviruses. Microorganisms, 9(8), 1731.

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Cell Culture and Virus Propagation Protocol

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African Green Monkey Vero E6 (ATCC® CRL-1586™), MRC-5 (ATCC® CCL-171™), human embryonic kidney 293T cells (ATCC® CRL-3216™) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS, Life Technologies), while HCT-8 (ATCC® CCL-244) were cultured in RPMI supplemented with 10% FBS. Calu-3 cells (ATCC® HTB-55™) were cultured in EMEM supplemented with nonessential amino acids and 10% FBS. All cells were cultured in the presence of 100 U/mL penicillin and 100 μg/mL streptomycin (Life Technologies), were maintained at 37 °C in a humidiﬁed atmosphere supplemented with 5% CO2, and periodically tested for the absence of mycoplasma contamination.
SARS-CoV-2/NL/2020 strain was obtained from European Virus Archive Global (EVAg) and was propagated and titrated in Vero E6 cells. HCoV-OC43 (VR-1558™) and HCoV-229E (VR-740™) strains were purchased from ATCC and were propagated in HCT-8 or MRC-5 cells, respectively.

Mercorelli B., Desantis J., Celegato M., Bazzacco A., Siragusa L., Benedetti P., Eleuteri M., Croci F., Cruciani G., Goracci L, & Loregian A. (2022). Discovery of novel SARS-CoV-2 inhibitors targeting the main protease Mpro by virtual screenings and hit optimization. Antiviral Research, 204, 105350.

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Culturing Human Colorectal Cancer Cell Lines

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Human CRC cell lines HCT116, HCT8, HT29, SW620, SW480 and DLD-1, and corresponding normal colonic epithelial cell (FHC) were purchased from ATCC. These cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, USA) with 100 U/ml penicillin, 0.1 mg/ml streptomycin, and 10% fetal bovine serum (FBS, Gibco,USA) at 37 ℃ supplied with 5% CO₂ atmosphere.

Chen X., Zeng K., Xu M., Hu X., Liu X., Xu T., He B., Pan Y., Sun H, & Wang S. (2018). SP1-induced lncRNA-ZFAS1 contributes to colorectal cancer progression via the miR-150-5p/VEGFA axis. Cell Death & Disease, 9(10), 982.

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Cell Lines for STEC Adhesion Profiles

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The human laryngeal epithelioma cell line HEp-2 (ATCC CCL-23; American Type Culture Collection, USA), the human ileocecal carcinoma cell line HCT-8 (ATCC CCL-244), and the human colonic carcinoma cell line T84 (ATCC CCL-248) were used for investigating STEC adhesion profiles. The three cell lines were grown with DMEM supplemented with 10% fetal bovine serum (FBS; Lonza) and antibiotics (2% penicillin G-streptomycin–amphotericin B) at 37°C, in an atmosphere of 5% CO₂. The T84 cells were differentiated by growing five days in 24-well plates coated with collagen I (Roche). About 10⁶ viable cells were used for each bacterial infection in 24-well plates.

Pradel N., Etienne-Mesmin L., Thévenot J., Cordonnier C., Blanquet-Diot S, & Livrelli V. (2015). In vitro adhesion properties of Shiga toxin-producing Escherichia coli isolated from cattle, food, and humans. Frontiers in Microbiology, 6, 156.

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Culturing HCT-8 and Vero Cell Lines

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The human intestinal cell line HCT-8 (ATCC CCL-244, Manassas, VA, USA) was maintained in RPMI-1640 medium (ATCC) and the monkey kidney cell line Vero (ATCC CCL-81) was grown in DMEM/F12 (Sigma Aldrich, St. Louis, MO USA). Both media were supplemented with 10% fetal bovine serum (FBS, Internegocios S.A., Buenos Aires, Argentina), 100 U/ml penicillin and 100 μg/ml streptomycin. Additionally, 1mM L-glutamine, 10 mM sodium pyruvate, 10 mM HEPES, 10 mM glucose were also added to HCT-8 cultures. Cells were grown at 37°C in a humidified 5% CO₂ incubator. Cells were subcultured until 80% confluence was obtained (7-10 days) in antibiotic-free medium. For growth-arrested conditions, medium without FBS was used.

Garimano N., Scalise M.L., Gómez F., Amaral M.M, & Ibarra C. (2022). Intestinal mucus-derived metabolites modulate virulence of a clade 8 enterohemorrhagic Escherichia coli O157:H7. Frontiers in Cellular and Infection Microbiology, 12, 975173.

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Cell Line Cultivation for Colon Cancer Research

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Human colon cancer cell lines (HT-29, HCT-116, HCT-8, Caco-2, SW620) and normal human intestinal epithelial cell line (FHC) were provided by ATCC (Manassas, VA, USA). HT-29 and HCT-8 were cultured with Dulbecco’s modified eagle medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (FBS) (Gibco, Rockville, MD, USA), penicillin (100 U/mL), and streptomycin (100 ug/mL), while HCT-116, SW620 and Caco-2 were cultured with Roswell Park Memorial Institute 1640 (RPMI 1640) medium. All cells were cultured in a 37°C cell incubator with 5% CO₂.

Liu J., Shi Z., Ma Y., Fu L, & Yi M. (2020). MOB1 Inhibits Malignant Progression of Colorectal Cancer by Targeting PAK2. OncoTargets and therapy, 13, 8803-8811.

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Cultivation and Passaging of Human Colon Cell Lines

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Human colon cancer cell lines HT29, HCT-8, HCT-116, and normal human intestinal epithelial cell line FHC were obtained from ATCC (Manassas, VA, USA). Human colon cancer cell lines HT29 and HCT-8 were cultured in Dulbecco’s modified eagle medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, USA) high glucose medium containing 10% fetal bovine serum (FBS) (Gibco, Rockville, MD, USA), penicillin (100U/mL) and streptomycin (100ug/mL). HCT- 116 was cultured in Roswell Park Memorial Institute 1640 (RPMI 1640) medium containing 10% fetal bovine serum and antibiotics. All cells were cultured in an incubator at 37 °C and 5% CO2. When the cells grew to 80−90% confluence, they were passaged with 1 × trypsin + ethylene diamine tetraacetic acid (EDTA) digestion.

Liu N., Dou L, & Zhang X. (2020). LncRNA PTTG3P Sponge Absorbs microRNA-155-5P to Promote Metastasis of Colorectal Cancer. OncoTargets and therapy, 13, 5283-5291.

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Characterization of CRC Cell Lines

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The CRC cell lines SW403, Colo201, KM12, HCT116, HCT15, RW2982, RKO, Caco2, HCT8, HT29, SW620 and SW948 were obtained from the ATCC. The KRAS mutant HCT116 cell line and its isogenic derivative, Hke3 in which mutant KRAS has been deleted by homologous recombination have been previously described [15 (link)] was kindly provided by Dr. S.Shirasawa to Dr. L Klampfer. Non transformed mutant KRAS inducible rat intestinal epithelial cells (IEC-I Kras) were kindly provided by Dr.Raymond DuBois [32 (link)]. This cell line expresses Kras^Val12 under the control of the lac operon, and is induced by treatment with 5 mM Isopropyl β-D-1-thiogalactopyranoside (IPTG, Life Technologies, Inc., Gaithersburg, MD) [32 (link)]. The HCT116 p21−/− cell line along with the parental HCT 116 p21 +/+ was kindly provided by Prof. Bert Vogelstein. All cell lines were cultured in MEM (Gibco BRL), supplemented with 10% FBS, 2mM L-Glutamine and 1% penicillin streptomycin. For serum free experiments the cell culture media PC-1 without L- Gln was used (Lonza #77232).

Maitra R., Seetharam R., Tesfa L., Augustine T.A., Klampfer L., Coffey M.C., Mariadason J.M, & Goel S. (2014). Oncolytic reovirus preferentially induces apoptosis in KRAS mutant colorectal cancer cells, and synergizes with irinotecan. Oncotarget, 5(9), 2807-2819.

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Culturing Colon Cell Lines

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Colon cancer cell lines (SW480, SW620 and HCT8) and normal human colon epithelial cell line (HIEC and NCM460) were obtained from ATCC (American Typical Culture Center). All cells were cultured in DMEM medium supplemented with 10% fetal bovine serum (GIBCO), 100units/mL penicillin and 100μg/mL streptomycin in an incubator with 5% CO₂ at 37°C.

Dai Y., Wang M., Wu H., Xiao M., Liu H, & Zhang D. (2016). Loss of FOXN3 in colon cancer activates beta-catenin/TCF signaling and promotes the growth and migration of cancer cells. Oncotarget, 8(6), 9783-9793.

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Silencing ORMDL1 in Colorectal Cancer Cells

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All cell lines (HCT15, HCT8, SW480, SW620, LOVO, DLD1, SW48, HCT116) used in this study were obtained from ATCC. Cell lines were typically cultured at 37 °C in an incubator with 5% CO₂. The SiRNAs used were as follows: siORMDL1#1: 5′- GGAGTTGGCTTGCTTCATA-3′, and siORMDL1#2: 5′- CTGGCAAGTTTCTATACGA-3′. Negative scramble fragment (RiboBio Co., China) was used as a control.

Wang Q., Liu W., Chen S., Luo Q., Li Y., Peng S., Wang H., Liu X, & Chen D. (2021). ORMDL1 is upregulated and associated with favorable outcomes in colorectal cancer. Translational Oncology, 14(10), 101171.

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