t84 human colon epithelial cells  (ATCC)

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Silver nanoparticles at sub-cytotoxic levels increase enteric pathogen invasion by compromising intestinal epithelial barrier integrity

doi: 10.3389/fcimb.2026.1745955

Figure Lengend Snippet: Effect of 10 and 20 µg/mL AgNPs pretreatment on bacterial adhesion, invasion, and persistence in infected T84 epithelial cells. T84 cells were pretreated or 24 hours with 10 nm AgNPs at 10 or 20 µg/mL, followed infection with Salmonella enterica serovar Heidelberg strain 146. (a) Bacterial adhesion, invasion, and persistence were quantified by measuring CFU/mL (as described in M&Ms). (b) Bacterial survival percentage in T84 cells during adhesion, invasion and persistence following AgNPs treatment. Survival was calculated by dividing CFU/mL (as described in M&Ms) for each condition by its corresponding control (set to 100% and therefore not shown as a separate bar) and expressed as a percentage. Bars represent mean ± SD from three independent experiments performed in biological triplicates. Asterisks indicate statistically significant differences compared to respective untreated infection controls (** p < 0.005, *** p < 0.0005; unpaired t-test).

Article Snippet: Human colon carcinoma T84 epithelial cells (ATCC ® CCL-248TM) were cultured in complete medium composed of Dulbecco’s Modified Eagle Medium/Ham’s F-12 (DMEM/F-12) supplemented with l-glutamine and HEPES (ATCC, Manassas, VA, USA), 10% fetal bovine serum (FBS), 1% penicillin-streptomycin and 0.1% fungizone (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Infection, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Silver nanoparticles at sub-cytotoxic levels increase enteric pathogen invasion by compromising intestinal epithelial barrier integrity

doi: 10.3389/fcimb.2026.1745955

Figure Lengend Snippet: Relative expression of genes related to cell-cell junction and epithelial barrier function in T84 cells following AgNPs exposure and bacterial infection. T84 cells were pretreated with 10 nm AgNPs at either 10 µg/mL or 20 µg/mL, alone or in combination with S. enterica under invasion (1 h + gentamicin) or persistence (24 h + gentamicin) conditions. Invasion control consists of infected cells that are not pretreated with AgNPs but treated with gentamicin for 1 hour, while persistence control consists of infected cells not pretreated with AgNPs but treated with gentamicin for 24 hours. Gene expression (as described in M&Ms) was measured using RT² Profiler PCR arrays and is represented as Log 2 fold regulation relative to untreated control cells. (a) Focal adhesion genes, (b) Gap junction genes, (c) Tight junction genes, and (d) Adherens Junctions, desmosomes, and hemidesmosomes genes. Bars represent mean ± SE from biological triplicates. Asterisks indicate statistically significant differences compared to uninfected and AgNPs untreated control (p < 0.05) (*p < 0.05, **p < 0.005; unpaired t-test).

Article Snippet: Human colon carcinoma T84 epithelial cells (ATCC ® CCL-248TM) were cultured in complete medium composed of Dulbecco’s Modified Eagle Medium/Ham’s F-12 (DMEM/F-12) supplemented with l-glutamine and HEPES (ATCC, Manassas, VA, USA), 10% fetal bovine serum (FBS), 1% penicillin-streptomycin and 0.1% fungizone (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Expressing, Infection, Control, Gene Expression

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Silver nanoparticles at sub-cytotoxic levels increase enteric pathogen invasion by compromising intestinal epithelial barrier integrity

doi: 10.3389/fcimb.2026.1745955

Figure Lengend Snippet: Concentration of pro- and anti-inflammatory cytokines in T84 epithelial cells following AgNPs exposure and bacterial infection. Cytokine concentrations in the supernatants of T84 cells pretreated with 10 µg/mL or 20 µg/mL of 10 nm AgNPs and exposed to S. enterica under invasion or persistence conditions. (a) Pro-inflammatory cytokines and (b) Anti-inflammatory cytokine. Bars represent mean ± SE from biological triplicates. Asterisks indicate statistically significant differences compared to uninfected and AgNPs untreated control (p < 0.05).

Article Snippet: Human colon carcinoma T84 epithelial cells (ATCC ® CCL-248TM) were cultured in complete medium composed of Dulbecco’s Modified Eagle Medium/Ham’s F-12 (DMEM/F-12) supplemented with l-glutamine and HEPES (ATCC, Manassas, VA, USA), 10% fetal bovine serum (FBS), 1% penicillin-streptomycin and 0.1% fungizone (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Concentration Assay, Infection, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Silver nanoparticles at sub-cytotoxic levels increase enteric pathogen invasion by compromising intestinal epithelial barrier integrity

doi: 10.3389/fcimb.2026.1745955

Figure Lengend Snippet: Concentration of growth factors, chemokines, and Th2 cytokines in T84 epithelial cells following AgNPs exposure and bacterial infection. T84 cells were treated with 10 nm AgNPs at 10 µg/mL or 20 µg/mL, either alone or in combination with S. enterica invasion or persistence. Invasion control consisted of infected cells that were not pretreated with AgNPs but treated with gentamicin for 1 hour, while persistence control consisted of infected cells not pretreated with AgNPs but treated with gentamicin for 24 hours. (a) Growth factors, (b) chemokines, and (c) Th2 cytokines. Bars represent mean ± SE from biological triplicates. Asterisks indicate statistically significant differences compared to control (p < 0.05).

Article Snippet: Human colon carcinoma T84 epithelial cells (ATCC ® CCL-248TM) were cultured in complete medium composed of Dulbecco’s Modified Eagle Medium/Ham’s F-12 (DMEM/F-12) supplemented with l-glutamine and HEPES (ATCC, Manassas, VA, USA), 10% fetal bovine serum (FBS), 1% penicillin-streptomycin and 0.1% fungizone (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Concentration Assay, Infection, Control

Journal: Infection and Immunity

Article Title: ETEC biofilms are regulated by magnesium and lactate bioavailability

doi: 10.1128/iai.00243-25

Figure Lengend Snippet: Adherence of ETEC H10407 biofilms to epithelial cells. ETEC H10407 was grown in 4AA-lactate with 10 mM magnesium chloride to induce biofilm formation or 4AA-lactate with 0.25 mM magnesium chloride for planktonic conditions. ETEC H10407 biofilms and planktonic ETEC H10407 were resuspended and applied to T84 cells at 10 6 CFUs per well. Wells were then washed to remove unattached bacteria, and remaining cells and bacteria were resuspended and plated for CFUs ( A ). CD1 mice were given streptomycin to clear their normal flora and then orally infected with 10 8 CFU ETEC. Twenty-four hours post-infection, the jejunal tissue of the mice was harvested and processed to plate for CFUs ( B ). T84 data are compiled from three independent experiments. Mouse data are compiled from two independent experiments in which a total of nine mice were dosed with low magnesium cultured ETEC H10407 and seven were dosed with 4AA-lactate 10 mM magnesium chloride biofilm ETEC H10407 . Data were analyzed by t -test; *, P < 0.05; ***, P < 0.001.

Article Snippet: T84 human colonic epithelial cells (CCL-248) were purchased from ATCC and cultured on 24-well tissue culture-treated plates in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 5% fetal bovine serum.

Techniques: Bacteria, Infection, Cell Culture

Journal: Archives of toxicology

Article Title: Urolithin A attenuates arsenic-induced gut barrier dysfunction

doi: 10.1007/s00204-022-03232-2

Figure Lengend Snippet: UroA protects against iAs3+ induced cytotoxicity in colon epithelial cells. T84 cells (2 × 104) per well were grown O/N in 96 well plate. Next day, cells were treated with iAs3+ (0.1, 0.3, 1, 3, 10, 30, 100 μM) in the presence of UroA (25 μM) or vehicle (DMSO, 0.05%) for (A) 24 h, (B) 48 h and (C) 72 h. Cell viability was determined using alamarBlue assay. Percent control cell viability against iAs3+ dose were plotted (log(inhibitor) vs. normalized response (variable slope) curve fit). **p < 0.01, ***p < 0.001, Two-way Repeated Measures ANOVA with Geisser-Greenhouse’s Correction followed by Tukey’s Multiple Comparisons Test between iAs3+ and iAs3++UroA. Each point in each curve represents the mean ± SD from three independent experiments.

Article Snippet: The human colon epithelial carcinoma cell line T84 (ATCC # CCL-248 TM )) was maintained in DMEM: F-12 Medium (Cytiva # SH30261.01), supplemented with 10% fetal bovine serum, 1X penicillin-streptomycin solution (100 U/ml penicillin, and 100 μg/ml streptomycin (Sigma Aldrich) in a humidified atmosphere (at 37 °C in a 5% CO 2 incubator).

Techniques: Alamar Blue Assay, Control

Journal: Archives of toxicology

Article Title: Urolithin A attenuates arsenic-induced gut barrier dysfunction

doi: 10.1007/s00204-022-03232-2

Figure Lengend Snippet: (A) T84 cells were treated with iAs+3 (0, 1, 5, 10, 50, 100 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) for 48 h. Bar graph and images representing apoptosis in T84 cells after the indicated treatment estimated by Annexin V/PI assay. (B) Representative fluorescence image of JC1 stained T84 cells representing the effect of iAs+3 in presence of vehicle or UroA on mitochondrial permeability (red to the green shift of fluorescence). Bar graphs showing the detection of JC-1 fluorescence as J aggregates (red) vs J monomers (green). Results are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, Statistics performed using 2way ANOVA in GraphPad Prism software. Error bar, mean ± SEM (n=4–5)

Article Snippet: The human colon epithelial carcinoma cell line T84 (ATCC # CCL-248 TM )) was maintained in DMEM: F-12 Medium (Cytiva # SH30261.01), supplemented with 10% fetal bovine serum, 1X penicillin-streptomycin solution (100 U/ml penicillin, and 100 μg/ml streptomycin (Sigma Aldrich) in a humidified atmosphere (at 37 °C in a 5% CO 2 incubator).

Techniques: Fluorescence, Staining, Permeability, Software

Journal: Archives of toxicology

Article Title: Urolithin A attenuates arsenic-induced gut barrier dysfunction

doi: 10.1007/s00204-022-03232-2

Figure Lengend Snippet: (A) T84 cells were treated with iAs+3 (0, 0.05, 0.1, 1, 5, 10, 50, 100 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) for 12 h. Representative fluorescence image and bar graph are showing ROS generation as green fluorescence from DCFDA stained T84 cells. (B) LDH release after 24 h incubation of T84 cells with iAs+3 (5, 10, 20 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) (C). T-84 cells were treated with UroA (10, 25 μM). T84 cells were treated with iAs+3 (10 μM) and different concentration of UroA (0, 0.01, 0.1, 1, 5, 10, 25 μM) for 24 h. The levels of GSH and GSSG were measured and the GSH/GSSG ratio was calculated. Untreated (UT) cells were used as control (100%). Results are representative of three independent experiments. Statistics performed using 2way ANOVA in GraphPad Prism software. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Error bar, mean ± SEM (n=4–8)

Article Snippet: The human colon epithelial carcinoma cell line T84 (ATCC # CCL-248 TM )) was maintained in DMEM: F-12 Medium (Cytiva # SH30261.01), supplemented with 10% fetal bovine serum, 1X penicillin-streptomycin solution (100 U/ml penicillin, and 100 μg/ml streptomycin (Sigma Aldrich) in a humidified atmosphere (at 37 °C in a 5% CO 2 incubator).

Techniques: Fluorescence, Staining, Incubation, Concentration Assay, Control, Software

Journal: Archives of toxicology

Article Title: Urolithin A attenuates arsenic-induced gut barrier dysfunction

doi: 10.1007/s00204-022-03232-2

Figure Lengend Snippet: (A) Schematic representation of in vitro permeability study with T84 monolayers (B) and (C) Monolayer T84 cells on transmembrane were treated with iAs+3 (0, 1, 5, 10, 20 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) for 24 h. FITC-dextran was added to these cells (top of the membrane) and incubated for 2 h at 37°C and FITC-dextran levels in the bottom chamber well was measured. TEER values were also measured. (B) LDH release after 24 h incubation of T84 monolayer cells with iAs+3 (5, 10, 20 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) (C) T84 monolayer cells were incubated with iAs+3 (5, 10 μM) without or with UroA (0, 10, 25 μM) for 24 h. IL-8 levels in supernatants were measured. *p < 0.05, **p < 0.01, ***p < 0.001, Statistics performed using 2way ANOVA in GraphPad Prism software. Error bar, mean ± SEM (n=4).

Article Snippet: The human colon epithelial carcinoma cell line T84 (ATCC # CCL-248 TM )) was maintained in DMEM: F-12 Medium (Cytiva # SH30261.01), supplemented with 10% fetal bovine serum, 1X penicillin-streptomycin solution (100 U/ml penicillin, and 100 μg/ml streptomycin (Sigma Aldrich) in a humidified atmosphere (at 37 °C in a 5% CO 2 incubator).

Techniques: In Vitro, Permeability, Membrane, Incubation, Software

Journal: Archives of toxicology

Article Title: Urolithin A attenuates arsenic-induced gut barrier dysfunction

doi: 10.1007/s00204-022-03232-2

Figure Lengend Snippet: T84 cells were treated with iAs+3 (0, 5, 10 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) for 24 h. (A) Protein expression of Zona occludens 1 (ZO1), claudin 4 (Cldn4) and occludin (Ocln) in T84 cells were determined by immunoblots. (B) The fold changes in mRNA levels of ZO1, Cldn4, Ocln, and ZO1 in T84 cells were determined by RT PCR method. (C) T84 cells were grown on chambered slides and treated with iAs+3 (10 μM) in presence of vehicle (DMSO-0.01%) or UroA (25 μM) for 24 h. The cells were stained with rabbit anti ZO-1, rabbit anti Ocln and mouse anti-Cldn4, followed by secondary antibody tagged with anti-rabbit Alexa 488 for ZO-1, Ocln and anti-mouse Alexa-594 for cldn-4. Nucleus was stained using DAPI. The confocal images were captured. Scale bars for T84 cells indicate 20 μm respectively. The fluorescence intensity (n = 15–20 cell membrane regions) was measured. Results are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, Statistics performed using one-way ANOVA in GraphPad Prism software. Error bar, mean ± SEM (n=3–5).

Article Snippet: The human colon epithelial carcinoma cell line T84 (ATCC # CCL-248 TM )) was maintained in DMEM: F-12 Medium (Cytiva # SH30261.01), supplemented with 10% fetal bovine serum, 1X penicillin-streptomycin solution (100 U/ml penicillin, and 100 μg/ml streptomycin (Sigma Aldrich) in a humidified atmosphere (at 37 °C in a 5% CO 2 incubator).

Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Staining, Fluorescence, Membrane, Software

Journal: Cells

Article Title: Heat-Stable Enterotoxin Secretions Assessed via ICP-MS Reveal Iron-Mediated Regulation of Virulence in CFA/I- and CS6-Expressing ETEC Isolates.

doi: 10.3390/cells12040567

Figure Lengend Snippet: Figure 2. ST-induced depletion of luminal iron alters ST activity in ETEC H10407 and 214-4. ST intoxication causes a sustained decrease and flux in luminal iron (Fe) that begins during peak secretion (30 min of intoxication) and extends beyond 180 min (A). Iron limitation via deferoxamine mesylate (100 µM) increases expression of fepA, cfaB, and cfaE in ETEC H10407 (B). ETEC H10407 expresses more CFA/I when cultured with deferoxamine mesylate (100 µM) in 4AA media (C). Iron limitation via deferoxamine mesylate (100 µM) increases expression of fepA but not cssA and cssB in ETEC 214-4 (D). ETEC 214-4 expresses more CS6 when cultured with exogenous ferrous ammonium sulfate (100 µM) in 4AA media (E). ST production and secretion in ETEC H10407 cultured in 4AA media are unchanged in the absence and presence of increasing amounts of deferoxamine mesylate (1.0, 10, 100 µM) (F). No significant differences were observed when supernatants from ETEC H10407 cultured in 4AA media in the absence and presence of increasing amounts of deferoxamine mesylate (1.0, 10, 100 µM) were applied to T84 cells for ST-mediated induction of cGMP (G). ST production and secretion in ETEC 214-4 are suppressed when cultured in 4AA media containing deferoxamine mesylate (100 µM) (H). ST activity from ETEC 214-4 is significantly reduced when cultured in 4AA media containing deferoxamine mesylate (1.0, 10, 100 µM) (I). Data were analyzed via one-way ANOVA with matched conditions. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: Human T84 colonic epithelial cells (ATCC CCL-248) were cultured regularly in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 5% fetal bovine serum (FBS).

Techniques: Activity Assay, Expressing, Cell Culture

Journal: Cells

Article Title: Heat-Stable Enterotoxin Secretions Assessed via ICP-MS Reveal Iron-Mediated Regulation of Virulence in CFA/I- and CS6-Expressing ETEC Isolates.

doi: 10.3390/cells12040567

Figure Lengend Snippet: Figure 3. ST production and activity are decreased in CFA/I+ ST-ETEC isolates cultured with deferoxamine, while changes in CFA/I production are isolate-specific. Select ST-only CFA/I+-ETEC isolates from GEMS were cultured in chemically defined 4AA media with or without the addition of deferoxamine mesylate (100 µM) to induce iron limitation overnight before ST production (A), which was measured using the ST competitive ELISA. Broadly, addition of deferoxamine decreases ST production in ETEC supernatants. ETEC isolates were then down-selected based on ST production threshold of 2.0 ng/µg total protein, and supernatants (10 µg) were applied to T84 cells to determine ST-mediated induction of cGMP (B). Again, deferoxamine decreases ST activity measured from ETEC supernatants. Cell lysates from the same cultures demonstrate variable CFA/I production in the presence or absence of deferoxamine (C). The represented data in (A,B) represent the means of two individual experiments. The data represented in (C) was confirmed two times. Data were analyzed via T-tests within individual strains between culture conditions *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: Human T84 colonic epithelial cells (ATCC CCL-248) were cultured regularly in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 5% fetal bovine serum (FBS).

Techniques: Activity Assay, Cell Culture, Competitive ELISA

Journal: Cells

Article Title: Heat-Stable Enterotoxin Secretions Assessed via ICP-MS Reveal Iron-Mediated Regulation of Virulence in CFA/I- and CS6-Expressing ETEC Isolates.

doi: 10.3390/cells12040567

Figure Lengend Snippet: Figure 4. ST production and activity are decreased in CS6+ ST-ETEC isolates cultured with deferox- amine, while changes in CS6 production are isolate-specific. Select ST-only CS6+-ETEC isolates from GEMS were cultured in chemically defined 4AA media with or without the addition of deferoxamine mesylate (100 µM) to induce iron limitation overnight before ST production (A) was measured using the ST competitive ELISA. Broadly, addition of deferoxamine decreases ST production in CS6+-ETEC supernatants. ETEC isolates were then down-selected based on ST production threshold of 2.0 ng/µg total protein, and supernatants (10 µg) were applied to T84 cells to determine ST-mediated induction of cGMP (B). Again, deferoxamine decreases ST activity measured from ETEC supernatants. ETEC 214-4 was grown in chemically defined 4AA media with or without deferoxamine mesylate (100 µM) to mid-log phase (OD 0.5) before RNA isolation using the hot phenol/chloroform method. RNA was reverse transcribed into cDNA, which was used for qPCR to determine relative transcripts of genes expressing ST (porcine variant, STp, estP) as compared to genes expressing 16S rRNA. These qPCR data confirm that iron limitation via deferoxamine suppresses ST expression in ETEC 214-4 (C). Cell lysates from CS6+-ETEC isolates demonstrate variable CS6 production in the presence or absence of deferoxamine (D). The represented data in (A,B) represent the means of two individual experiments. The data represented in (D) was confirmed two times. Data in panels A and B were analyzed via T-tests within individual strains between culture conditions. Data in panel C were analyzed via a one-sample T-test against a hypothetical mean of 1. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Article Snippet: Human T84 colonic epithelial cells (ATCC CCL-248) were cultured regularly in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 5% fetal bovine serum (FBS).

Techniques: Activity Assay, Cell Culture, Competitive ELISA, Isolation, Reverse Transcription, Expressing, Variant Assay