Caco-2 cells (ECACC, Salisbury, UK) were grown in MEM with Earle’s salts (PAA Laboratories GmbH, Pasching, Austria) containing 15% fetal bovine serum. HCT-116 cells (DSMZ, Braunschweig, Germany) were grown in McCoy’s 5A medium (Biowest, Nuaillé, France) containing 10% fetal bovine serum (PAA Laboratories). HCT-116 cells harbour an activating mutation in KRAS (G13D), while Caco-2 cells are KRAS wildtype. Culture media were supplemented with 2 mM glutamine, 1% non-essential amino acids, penicillin (100 U/ml), streptomycin (100 μg/ml) and amphotericin B (2.5 μg/ml). Cells were maintained in a humidified atmosphere at 37°C and 5% CO2. After cultures became 80% confluent (usually after 3 days), cells were trypsinized, centrifuged, and suspended in fresh medium. All cells used for experiments displayed > 95% viability and were seeded in 96-well plates, 6-well culture plates, 60-mm culture plates or ultra low-attachment plates (Corning Inc, Lowell, MA, USA). All experiments were carried out in duplicate and repeated at least three times.
Hct 116 cells
Manufactured by Leibniz Institute DSMZ
Sourced in Germany
HCT-116 cells are a human colon cancer cell line. They are commonly used in research to study various aspects of cancer biology and drug development.
Automatically generated - may contain errors
Lab products found in correlation
Mccoy s 5a medium, by Biowest (3 mentions)
Fetal bovine serum (fbs), by GE Healthcare (3 mentions)
60 mm culture plates, by Corning (2 mentions)
Earle s salts, by GE Healthcare (2 mentions)
Ultra low attachment plate, by Corning (2 mentions)
6 well culture plate, by Corning (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Mem medium, by Biowest (1 mentions)
Mem non essential amino acids, by Biowest (1 mentions)
Zell shield antibiotics, by Minerva Biolabs (1 mentions)
Glutamine, by Biowest (1 mentions)
Minimal essential medium, by Merck Group (1 mentions)
Mccoy s 5a modified medium, by Thermo Fisher Scientific (1 mentions)
Human caco 2 cells, by Leibniz Institute DSMZ (1 mentions)
Fetal bovine serum (fbs), by Bio&Sell (1 mentions)
Dmem high glucose medium, by Capricorn (1 mentions)
Dld 1 cells, by American Type Culture Collection (1 mentions)
4 protocols using hct 116 cells
1
Culturing Colorectal Cancer Cell Lines
2
Cell Culture Conditions for HCT-116 and Caco-2
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HCT-116 cells (DSMZ, Braunschweig, Germany) were grown in McCoy’s 5A medium (Biowest) containing 10% fetal bovine serum (FBS) (PAA Laboratories, Pasching, Austria). Caco-2 cells (ECACC, Salisbury, UK) were grown in MEM medium (Biowest, Nuaillé, France) containing 15% FBS (PAA Laboratories) and MEM non-essential amino acids (Biowest). Culture media were supplemented with 2 mM glutamine (Biowest) and Zell Shield antibiotics (Minerva Biolabs, Berlin, Germany). Cells were maintained in a humidified atmosphere at 37 °C and 5 °C CO2.
3
Caco-2 and HCT-116 Cells with E. coli Strains
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Human Caco-2 cells (DSMZ, Braunschweig, Germany) were cultured using minimal essential medium (Sigma-Aldrich, Taufkirchen, Germany) supplemented with 10% fetal bovine serum (Bio&Sell, Nürnberg, Germany) without supplementation of antibiotics. For culture of HCT-116 cells (DSMZ, Braunschweig, Germany), McCoy’s 5A modified medium (Thermo Fisher Scientific, Waltham, Massachusetts, USA) was supplemented with 10% fetal bovine serum without antibiotics. Ten different E. coli strains were isolated from ascitic fluid of patients with SBP and serotyping was performed by Robert Koch Institute (Berlin, Germany). For assay development, verification and validation, E. coli ATCC25922 (O6:Hnt) (ATCC, Manassas, Virginia, USA) was used. In addition, a P. mirabilis strain was extracted from ascites of a patient with SBP (for antibiograms, see online supplemental table 6 ). Bacteria were grown in Luria-Bertani broth at 37°C under agitation. Co-culture experiments were performed with live bacteria at multiplicities of infection (MOI) 0–10, supernatant of bacterial overnight culture (SN) or heat-inactivated (HI) bacteria (65°C, 5 min). For transwell experiments, see the Supplementary material and methods section.
4
Culturing of Colorectal Cancer Cell Lines
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HT-29 and HCT-116 cells (DSMZ, Braunschweig, Germany) were grown in McCoy's 5A medium (Biowest, Nuaillé, France) containing 10% fetal bovine serum (PAA Laboratories, Pasching, Austria). Caco-2 cells (ECACC, Salisbury, UK) were grown in MEM with Earle's salts (PAA Laboratories GmbH) containing 15% fetal bovine serum. DLD-1 cells (ATCC, US ) were grown in D-MEM high glucose medium (Capricorn scientific, GmbH), containing 15% fetal bovine serum. Culture media were supplemented with 2 mM glutamine, 1% non-essential amino acids, penicillin (100 U/ml), streptomycin (100 μg/ml) and amphotericin B (2.5 μg/ml) and cells were cultured in a humidified atmosphere at 37°C and 5% CO2. in 96-well plates, 6-well culture plates, 60-mm culture plates or ultra low-attachment plates (Corning Inc, Lowell, MA, USA). All experiments were carried out in duplicate and repeated at least three times.
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