The HT-29 cell line was purchased from the Cell Bank, Pasteur Institute of Iran. The HT-29 cell line is a human colorectal epithelial cell line that expresses TLR4. The cell line was used to test the bioactivity of purified FimH and the fusion protein MrpH. FimH. The TLR4 activity of the proteins was evaluated based on the induction of IL-8. Briefly, HT-29 cells were cultured in 24-well plates (Greiner, Germany) at a density of 5 × 104 cells/well in 1 ml fresh Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (Biosera, North America) and antibiotics (Biosera, North America). After overnight incubation, the cells were incubated for 5 h with 10 μg/ml of sterilized of FimH and fusion MrpH. FimH. Then, supernatants were collected and the expression of IL-8 was evaluated using the enzyme-linked immunosorbent assay (ELISA) (R and D systems, USA).
Ht 29
Manufactured by Greiner
Sourced in Germany
The HT-29 is a laboratory cell line derived from a human colorectal adenocarcinoma. It is commonly used in research as a model for studying various aspects of intestinal cell biology and disease processes.
Automatically generated - may contain errors
Lab products found in correlation
24 well plate, by Greiner (2 mentions)
Elisa, by R&D Systems (1 mentions)
Fetal bovine serum (fbs), by Biosera (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 rpmi, by Thermo Fisher Scientific (1 mentions)
Caco 2, by Leibniz Institute DSMZ (1 mentions)
Ht 29, by Leibniz Institute DSMZ (1 mentions)
Caco 2, by Greiner (1 mentions)
2 protocols using ht 29
1
Evaluation of FimH and MrpH Bioactivity in HT-29 Cells
2
Colon Cancer Cell Line Experiments
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The colon cancer cell lines, HT-29 and CACO-2, were purchased from DSMZ (Braunschweig, Germany). Each cell line was cultured in RPMI-1640 (RPMI; Life Technologies, Carlsbad, CA, USA) supplemented with 2 mM L-glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin (Life Technologies) with 10% heat-inactivated fetal calf serum (FCS) (Thermo Fisher Scientific, Inc., Waltham, MA, USA) and 10 mM HEPES. Both cell lines were grown at 37°C in a humidified atmosphere and 5% CO2.
For all experiments, 29,000 HT-29 cells/well or 19,000 CACO-2 cells/well were seeded onto 24-well plates (Greiner Bio-One GmbH, Frickenhausen, Germany) in 0.5 ml RPMI 10% FCS plus 10 mM HEPES and cultured for 3 days. Thereafter, cells were treated with M1 or M2 macrophage CM (for preparation see below) or 5-FU, alone or in combination, according to the schedule shown inFig. 1 .
For all experiments, 29,000 HT-29 cells/well or 19,000 CACO-2 cells/well were seeded onto 24-well plates (Greiner Bio-One GmbH, Frickenhausen, Germany) in 0.5 ml RPMI 10% FCS plus 10 mM HEPES and cultured for 3 days. Thereafter, cells were treated with M1 or M2 macrophage CM (for preparation see below) or 5-FU, alone or in combination, according to the schedule shown in
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