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.
Human caco 2 cells
Manufactured by Leibniz Institute DSMZ
Sourced in Germany
Caco-2 cells are a human epithelial colorectal adenocarcinoma cell line derived from the colon. They are commonly used in in vitro studies to evaluate drug permeability and intestinal absorption.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Minimal essential medium, by Merck Group (1 mentions)
Mccoy s 5a modified medium, by Thermo Fisher Scientific (1 mentions)
Hct 116 cells, by Leibniz Institute DSMZ (1 mentions)
Fetal bovine serum (fbs), by Bio&Sell (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
2 mercaptoethanol, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Harvard Bioscience (1 mentions)
Streptomycin, by Harvard Bioscience (1 mentions)
Mito serum extender, by Corning (1 mentions)
Penicillin, by Corning (1 mentions)
Streptomycin, by Corning (1 mentions)
3 protocols using human caco 2 cells
1
Caco-2 and HCT-116 Cells with E. coli Strains
2
Culturing Insulin Receptor-Expressing CHO-K1 and Caco-2 Cells
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CHO-K1 cells stably expressing human insulin receptor (hIR) and GLUT4-myc-GFP were a kind gift from Manoj K. Bhat (National Centre for Cell Science, University of Pune, India). Cells were maintained in Ham’s F12 culture medium supplemented with 100 μg/mL penicillin, 100 μg/mL streptomycin, 1% G418 and 10% fetal bovine serum (FBS) (all Life Technologies, Carlsbad, CA, USA). Human Caco-2 cells were obtained from DSMZ (Braunschweig, Germany) and maintained in MEM with Earle’s salts supplemented with 10% FBS, 100 μg/mL penicillin, 100 µg/mL streptomycin, and 0.1% 2-mercaptoethanol (all Life Technologies, Carlsbad, CA, USA). The cells were grown at 37 °C in a humidified atmosphere with 5% CO2.
3
Caco-2 Cell Culture and Barrier Integrity
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Human Caco-2 cells (DSMZ, Braunschweig, Germany) were maintained in MEM with Earle’s salts supplemented with 10% FBS and 100 U/mL penicillin/100 µg/mL streptomycin (Biochrom GmbH, Berlin, Germany) and grown at 37 °C in a humidified atmosphere (≥95%) with 5% CO2. For qPCR experiments, the cells were seeded in 12-well plates at 1.2 × 106 cells per well. For the TEER measurements in transwell inserts (8.4 mm, collagen-treated, 0.4 µm pore diameter), the cells were seeded at 1.65 × 105 per insert to reach confluency on the next day (Greiner Bio-One International GmbH, Kremsmünster, Austria). The cells were further maintained in Entero-STIM Intestinal Epithelium Differentiation Medium supplemented with 0.1% MITO+ Serum Extender (Corning, Wiesbaden, Germany) and 100 U/mL penicillin/100 µg/mL streptomycin, and the medium was changed daily. The qPCR experiment was carried out on day 5, when the cells were completely differentiated. The measurement of intestinal barrier integrity was performed on day 7, when TEER reached values of at least 500 Ω.
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