Ht 29

Manufactured by Merck Group

Sourced in United States, Germany, Italy, Macao, Poland

HT-29 is a cell line derived from a human colorectal adenocarcinoma. It is commonly used in cancer research for the study of colon cancer.

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Lab products found in correlation

Hct116, by Merck Group (33 mentions) Mccoy s 5a medium, by Merck Group (23 mentions) Hct116, by American Type Culture Collection (22 mentions) Ht 29, by American Type Culture Collection (17 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (17 mentions) Penicillin, by Merck Group (16 mentions) Streptomycin, by Merck Group (13 mentions) Rpmi 1640, by Merck Group (12 mentions) Fetal bovine serum (fbs), by Merck Group (12 mentions) Mcf 7, by Merck Group (11 mentions) Dulbecco modified eagle medium (dmem), by Merck Group (11 mentions) Caco 2, by Merck Group (11 mentions) Sw480, by Merck Group (10 mentions) Sw620, by Merck Group (10 mentions) Rpmi 1640 medium, by Merck Group (9 mentions) Rpmi 1640, by Thermo Fisher Scientific (9 mentions) Mcf 7, by American Type Culture Collection (9 mentions) Sw620, by American Type Culture Collection (9 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (8 mentions) Dld 1, by Merck Group (7 mentions)

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HT-29 cell line (2 citations)

111 protocols using ht 29

Effects of Pectin on Colon Cancer Cells

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The human colon carcinoma cell line, HT29 (ATCC, Manassas, VA, USA) were cultured in McCoy’s 5A medium (ATCC) containing 16.7 mmol/L glucose, 200 U/mL penicillin, 200 μg/mL streptomycin, and 10% fetal bovine serum (FBS) at 37°C in a humidified atmosphere of 5% CO₂. Cells were sub-cultured every 2 to 3 days. Before the start of experiments, HT29 cells were synchronized by incubation with 0.5% fetal calf serum (FCS) for 24 h.
All experiments were performed under proliferating conditions by incubating HT29 cells for 24 h at 37°C in the humidified atmosphere of 5% CO₂ in a sub-confluent state (75% confluence and with 10% FBS), in both the absence and presence of pectin (P7536, Sigma-Aldrich Co., St. Louis, MO, USA). This pectin was originally extracted with hot acidic water. Pectin was prepared by diluting 0.06 mg pectin powder in 1 mL McCoy’s 5A medium (309 μmol/L final concentration) and heating at 37°C for 30 min. HT29 cells were incubated with pectin for 24 h at 37°C.
At the end of the incubation period, HT29 cells were lysed in a buffer containing 20 mmol/L Tris-HCl, 137 mmol/L NaCl, 10% glycerol, 1% Nonidet-P40, 1 mmol/L phenylmethylsulfonyl fluoride, 10 mmol/L ethylenediaminetetraacetic acid, and 2 mmol/L sodium orthovanadate, and were immediately frozen at −80°C until molecular parameters were determined.

Zamorano-León J.J., Ballesteros S., de las Heras N., Alvarez-Sala L., de la Serna-Soto M., Zekri-Nechar K., Freixer G., Calvo-Rico B., Yang Z., García-García J.M., Lahera V, & López-Farré A.J. (2019). Effect of Pectin on the Expression of Proteins Associated with Mitochondrial Biogenesis and Cell Senescence in HT29-Human Colorectal Adenocarcinoma Cells. Preventive Nutrition and Food Science, 24(2), 187-196.

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Differentiation of HT-29 Cells into Goblet Cells

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Caco-2 intestinal epithelial cells (cat#: HTB-37) and HT-29 cells (cat#: HTB-38) were purchased from the American Type Culture Collection (Manassas, VA, USA). The cells were seeded at a concentration of 5 × 10⁵ in T-75 flasks in DMEM-based CGM. The medium was changed every 2 days, and the cells were subcultured when they reached 80% confluency. To induce the differentiation of HT-29 cells into mucus-secreting goblet cells (HT-29-MTX), HT-29 cells were seeded at a concentration of 5 × 10⁵ in T-75 flasks for 5 days, incubated with 0.1 µM MTX (Sigma-Aldrich; cat#: SIA-M9929) for 28 days, and cultured in DMEM-based CGM without MTX for 48 h [21 (link)]. The medium was replaced every 2 days. After differentiation, HT-29-MTX cell lines were harvested and stored in liquid nitrogen until use. For cell proliferation investigations, the HT-29-MTX cells were cultured in CGM at a concentration of 5 × 10⁵ cells in T-75 flasks. After 4–5 days, 80% confluence was achieved.

Phuangbubpha P., Thara S., Sriboonaied P., Saetan P., Tumnoi W, & Charoenpanich A. (2023). Optimizing THP-1 Macrophage Culture for an Immune-Responsive Human Intestinal Model. Cells, 12(10), 1427.

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Anticancer Effects of 5-FU, OXP, and IRI on CRC Cell Lines

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Human colorectal cancer (CRC) cell lines HCT-116 and HT-29 were obtained from the American Type Culture Collection and grown for 72 h in DMEM/F12 medium (Invitrogen, USA) containing 10% fetal bovine serum, 4 mM L-glutamine, 100 units/mL penicillin, and 0.1 mg/mL streptomycin (Invitrogen, USA) at 37 °С and 5% СО₂. HT-29 and HCT-116 cells were incubated for 72 h and 144 h in the medium that contained either 2 µM or 20 µM 5-FU, 2 µM or 20 µM OXP or 5 µM IRI (Sigma-Aldrich, USA). HT-29 cells were incubated for 72 h and 144 h in the medium that contained 2 µM or 20 µM 5-FU, 2 µM OXP or 5 µM IRI (Sigma-Aldrich, USA). HCT-116 cells were treated with OXP (2 µM or 20 µM OXP) for 72 h and 144 h.

Zinovieva O.L., Grineva E.N., Krasnov G.S., Karpov D.S., Zheltukhin A.O., Snezhkina A.V., Kudryavtseva A.V., Mashkova T.D, & Lisitsyn N.A. (2019). Treatment of cancer cells with chemotherapeutic drugs results in profound changes in expression of genes encoding aldehyde-metabolizing enzymes. Journal of Cancer, 10(18), 4256-4263.

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Culturing Human Cancer Cell Lines

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The human cancer cell lines breast adenocarcinoma (MCF-7), colon adenocarcinoma (Caco-2 and HT-29), and glioblastoma (U87MG) were obtained from the Sigma-Aldrich company (ECACC, Steinheim, Germany) and from the collection of the Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warsaw, Poland. All cell lines were cultured in DMEM media supplemented with heat-inactivated fetal bovine serum and antibiotics. Cells were passaged with 0.25% trypsin-EDTA after washing with phosphate-buffered saline. All cultures were carried out at 37 °C and in a CO₂ atmosphere (5%).

Pawłowska A.M., Żurek N., Kapusta I., De Leo M, & Braca A. (2023). Antioxidant and Antiproliferative Activities of Phenolic Extracts of Eriobotrya japonica (Thunb.) Lindl. Fruits and Leaves. Plants, 12(18), 3221.

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Cell Line Characterization for Cancer Research

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The following cell lines were used in the study: SW480, SW620, DLD-1, HT29, HCT116, LoVo (all human colon cancer), MC38 (mouse colon cancer), OE33 (human esophageal cancer), and HeLa (human cervical cancer). All the cell lines were purchased from American Type Culture Collection (ATCC), except for MC38 and OE33 which were purchased from Kerafast and Sigma, respectively. Early passage SW480, SW620, DLD-1, HT29, HCT116, and OE33 cell lines were cultured in Roswell Park Memorial Institute (RPMI-1640) media (R8758; Sigma). Early passage MC38, and HeLa cell lines were cultured in Dulbecco’s Modified Eagle Medium (DMEM) media (D2429; Sigma). All media were supplemented with 10% fetal bovine serum (FBS, 16000-044; Thermo Fisher), and 100 IU/ml penicillin/streptomycin (15140-122; Thermo Fisher). All cells were cultured under sterile conditions and maintained in a 37 °C incubator with 5% CO₂, regularly tested for mycoplasma and genetically authenticated by CRUK Cancer Centre Genomics Facility, Leeds, UK. Cell lines were passaged upon reaching ∼80–90% confluency and their morphology was regularly checked to ensure the absence of cross contamination between cell lines or mycoplasma contamination. To determine cell number, the automated cell counter NucleoCounter NC-100 (Chemometec) or manual hemocytometers were used.

Yuzhalin A.E., Gordon-Weeks A.N., Tognoli M.L., Jones K., Markelc B., Konietzny R., Fischer R., Muth A., O’Neill E., Thompson P.R., Venables P.J., Kessler B.M., Lim S.Y, & Muschel R.J. (2018). Colorectal cancer liver metastatic growth depends on PAD4-driven citrullination of the extracellular matrix. Nature Communications, 9, 4783.

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Characterization of Attenuated HSV-1 Strain

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The human CRC cell line WiDr and African green monkey kidney cell line Vero were obtained from American Type Culture Collection (Manassas, VA, USA). The human CRC cell line CW2 was obtained from the RIKEN Cell Bank (Tsukuba, Japan). The human CRC cell line HT-29 was kindly donated by Dr. Suguru Yamada (Nagoya University, Japan). HT-29, WiDr, CW2, and Vero cells were grown in DMEM (Sigma, Tokyo, Japan) supplemented with 10% fetal calf serum (FCS) and 1% penicillin/streptomycin (Gibco). C-REV is a highly attenuated mutant clone derived from HSV-1 strain HF.⁴⁵ (link) The virus was propagated in Vero cells and stored in aliquots at −80°C. C-REV was diluted in PBS for in vivo and in vitro experiments. Viral titers were assayed in Vero cells and expressed as plaque-forming units per milliliter (PFU/mL).

Wu Z., Ichinose T., Naoe Y., Matsumura S., Villalobos I.B., Eissa I.R., Yamada S., Miyajima N., Morimoto D., Mukoyama N., Nishikawa Y., Koide Y., Kodera Y., Tanaka M, & Kasuya H. (2019). Combination of Cetuximab and Oncolytic Virus Canerpaturev Synergistically Inhibits Human Colorectal Cancer Growth. Molecular Therapy Oncolytics, 13, 107-115.

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Probiotic Strains and Epithelial Cell Interaction

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Lactobacillus brevis KY21 (isolated from Korean infant feces), L. salivarius E4191 (isolated from Egyptian infant feces), and Leuconostoc mesenteroides subsp. mesenterides KDK411 (isolated from Kimchi) used in this study were generated in the Sae Hun Kim group (Food Microbiology Laboratory at Korea University, Korea). The strains were cultured in de Man, Rogosa, and Sharpe (MRS) broth (Difco, USA) at 37℃ for 18 h. The stock cultures were maintained at −80℃, using 50% glycerol as a cryoprotectant. The strains were sub-cultured three times prior to use.
The human epithelial cell line, HT-29, was obtained from the Korea Cell Line Bank (KCLB, Korea). The HT-29 cells were maintained at 37℃ with 5% CO₂ in RPMI 1640 medium (HyClone, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (HyClone) and 1% penicillin/streptomycin (P-S; HyClone). The HT-29 cells were seeded on a six-well plate (Palcon, USA) at a density of 1×10⁶ cells/well for western blot and confocal analyses. The HT-29 cells were pre-treated with 100 μg/mL of GS extract or 10⁹ CFU/mL probiotic bacteria for 2 h and cultured with 1 μg/mL of LPS (Escherichia coli strain, 0111:B4, Sigma, USA).

Kim Y., Koh J.H., Ahn Y.J., Oh S, & Kim S.H. (2015). The Synergic Anti-inflammatory Impact of Gleditsia sinensis Lam. and Lactobacillus brevis KY21 on Intestinal Epithelial Cells in a DSS-induced Colitis Model. Korean Journal for Food Science of Animal Resources, 35(5), 604-610.

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Cell Culture of Human Cancer Cell Lines

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Human colon adenocarcinoma cell line HT-29 (Catalogue Number 91072201, ATCC HTB-38) and human breast carcinoma cell line ZR-75-1 (Catalogue Number 87012601, ATCC CRL 1500) were acquired from the European Collection of Authenticated Cell Cultures (ECCAC). In accordance with ECCAC instructions, HT-29 cells were cultured in McCoy’s 5a medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS) (Sigma Aldrich), 2 mM of L-Glutamine (Sigma-Aldrich), 100 U/ml of penicillin and 100 μg/ml of streptomycin (Sigma-Aldrich) at 37°C and 5% CO₂. Cells were passaged when 80–90% confluence was reached, and the media was changed every 2–3 days. In accordance with ECCAC instructions, ZR-75-1 cells were cultured in RPMI 1640 (Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich), 1 mM sodium pyruvate (Sigma-Aldrich), 2 mM of L-Glutamine (Sigma-Aldrich) and 100 U/ml of penicillin and 100 μg/ml of streptomycin (Sigma-Aldrich) at 37°C and 5% CO₂. Cells were passaged when 70–80% confluence was reached, and the media was changed every 2–3 days.

Salguero-Aranda C., Sancho-Mensat D., Canals-Lorente B., Sultan S., Reginald A, & Chapman L. (2019). STAT6 knockdown using multiple siRNA sequences inhibits proliferation and induces apoptosis of human colorectal and breast cancer cell lines. PLoS ONE, 14(5), e0207558.

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FGL1 Modulates Intestinal Inflammation

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The human colonic adenoma cell line HT-29 (ATCC, United States) was cultured with Dulbecco’s modified Eagle’s medium, supplemented with 10% fetal bovine serum, 100 μg/mL penicillin, and 100 U/mL streptomycin at 37 °C with 5% CO₂. The HT-29 cells were stimulated with 100 ng/mL lipopolysaccharide (LPS, Sigma, United States) to establish a cell model of intestinal inflammation. To uncover the impact of FGL1 on intestinal inflammation, the HT-29 cells were transfected with FGL1 siRNA and plasmid DNA (Nanjing KeyGen Biotech Co., Ltd., China) before stimulation with LPS. The transfection efficiency was determined by examining the mRNA expression of FGL1.

Sun X.L., Qiao L.C., Gong J., Wen K., Xu Z.Z, & Yang B.L. (2021). Proteomics identifies a novel role of fibrinogen-like protein 1 in Crohn’s disease. World Journal of Gastroenterology, 27(35), 5946-5957.

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Cytotoxicity Evaluation of Capsaicin on Intestinal Cells

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Because of the known cytotoxicity of capsaicin [12 (link), 13 (link)] (a main substance responsible for the hot and spicy taste in the chili), capsaicin (305.41 g/mol molecular weight; Sigma-Aldrich, St. Louis, MO, USA) were tested with enterocytes (Caco-2 and HT-29 cell lines). As such, the human colorectal adenocarcinoma cells, Caco-2 (ATCC HTB-37) and HT-29 (ATCC HTB-38), from the American Type Culture Collection (Manassas, VA, USA) were maintained in supplemented Dulbecco’s modified Eagle medium (DMEM) and McCoy’s 5a modified medium, respectively, at 37°C under 5% CO₂ and sub-cultured before use in the experiments. Then, capsaicin in the different concentrations (0.02, 0.2 and 2 mM) was incubated with the enterocytes for 24 h before the determination of cell viability using tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) solution (Thermo Fisher Scientific, Rockford, IL, USA) [38 (link)]. The activated cells were incubated with 0.5 mg/mL of MTT solution for 2 h at 37°C in the dark and diluted by dimethyl sulfoxide (DMSO; Thermo Fisher Scientific) before measurement with a Varioskan Flash microplate reader at absorbance of optical density (OD) 570 nm.

Panpetch W., Visitchanakun P., Saisorn W., Sawatpanich A., Chatthanathon P., Somboonna N., Tumwasorn S, & Leelahavanichkul A. (2021). Lactobacillus rhamnosus attenuates Thai chili extracts induced gut inflammation and dysbiosis despite capsaicin bactericidal effect against the probiotics, a possible toxicity of high dose capsaicin. PLoS ONE, 16(12), e0261189.

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