epiintestinal 3d microtissues  (MatTek)

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) and (C) iAs+3 (0, 1, 5, 10, 20, 25, 50 μM) (B) and (D) iAs+3 (25 μM) with UroA (0, 5, 10, 25, 50 μM) were added to the apical compartment of EpiIntestinal 3D microtissues grown on Transwell inserts and incubated at 37 °C for 24 h and 48 h, respectively. FITC-dextran was added to these cells (apical compartment) and incubated for 2 h at 37°C and FITC-dextran levels in the bottom chamber well (basal compartment) was measured. *p < 0.05, **p < 0.01, ***p < 0.001, Statistics performed using one-way ANOVA in GraphPad Prism software. Error bar, mean ± SEM (n=4).

Article Snippet: EpiIntestinal 3D microtissues (SMI-100 and SMI-196) were obtained from MatTek Corporation (Ashland, MA, USA) and cultured according to the manufacturer’s instructions (24-well and 96-well format respectively) in a specially formulated culture medium (MatTek Corporation).

Techniques: 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: Microphotographs EpiIntestinal3D microtissues treated with iAs+3 (10 μM) in presence or absence of UroA (25μM) were stained with rabbit anti ZO-1 followed by secondary antibody tagged with anti-rabbit Alexa 488. Nucleus was stained using DAPI. The confocal images were captured. Scale bars indicate 50 μm. 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=4).

Article Snippet: EpiIntestinal 3D microtissues (SMI-100 and SMI-196) were obtained from MatTek Corporation (Ashland, MA, USA) and cultured according to the manufacturer’s instructions (24-well and 96-well format respectively) in a specially formulated culture medium (MatTek Corporation).

Techniques: Staining, Fluorescence, Membrane, 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: EpiIntestinal3D microtissues were treated with iAs+3 (5,10,20 μM) in presence or absence of UroA (25μM) for 24h. (A) LDH release (B) IL-8 levels and (C) TNFα 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: EpiIntestinal 3D microtissues (SMI-100 and SMI-196) were obtained from MatTek Corporation (Ashland, MA, USA) and cultured according to the manufacturer’s instructions (24-well and 96-well format respectively) in a specially formulated culture medium (MatTek Corporation).

Techniques: 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: For chronic exposure of iAs+3 in human intestinal organoids, iAs+3 (0, 1, 5 μM) with or without UroA (10 μM) were added to the apical compartment of EpiIntestinal 3D microtissues grown on Transwell inserts and incubated at 37 °C for 14 days. Fresh treatments were added every alternate days and supernatants were stored. (A) On 7th and 14th day, FITC-dextran was added to these cells (apical compartment) and incubated for 2 h at 37°C and FITC-dextran levels in the bottom chamber well (basal compartment) was measured. (B) EpiIntestinal 3D microtissues were treated with iAs3+ (1, 5 μM) in presence or absence of UroA (10μM) for 14 days. The supernatants were collected every 3 days for 14 days and pooled. LDH release in these cumulative supernatants at day 14 was measured. (C) IL-8 levels and (D) TNFα levels in supernatants were measured. *p < 0.05, **p < 0.01, ***p < 0.001, Statistics performed using one-way ANOVA in GraphPad Prism software. Error bar, mean ± SEM (n=4).

Article Snippet: EpiIntestinal 3D microtissues (SMI-100 and SMI-196) were obtained from MatTek Corporation (Ashland, MA, USA) and cultured according to the manufacturer’s instructions (24-well and 96-well format respectively) in a specially formulated culture medium (MatTek Corporation).

Techniques: 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: (A) and (C) iAs+3 (0, 1, 5, 10, 20, 25, 50 μM) (B) and (D) iAs+3 (25 μM) with UroA (0, 5, 10, 25, 50 μM) were added to the apical compartment of EpiIntestinal 3D microtissues grown on Transwell inserts and incubated at 37 °C for 24 h and 48 h, respectively. FITC-dextran was added to these cells (apical compartment) and incubated for 2 h at 37°C and FITC-dextran levels in the bottom chamber well (basal compartment) was measured. *p < 0.05, **p < 0.01, ***p < 0.001, Statistics performed using one-way ANOVA in GraphPad Prism software. Error bar, mean ± SEM (n=4).

Article Snippet: Primary human small intestinal epithelial cultures EpiIntestinal 3D microtissues (SMI-100 and SMI-196) were obtained from MatTek Corporation (Ashland, MA, USA) and cultured according to the manufacturer’s instructions (24-well and 96-well format respectively) in a specially formulated culture medium (MatTek Corporation).

Techniques: 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: For chronic exposure of iAs+3 in human intestinal organoids, iAs+3 (0, 1, 5 μM) with or without UroA (10 μM) were added to the apical compartment of EpiIntestinal 3D microtissues grown on Transwell inserts and incubated at 37 °C for 14 days. Fresh treatments were added every alternate days and supernatants were stored. (A) On 7th and 14th day, FITC-dextran was added to these cells (apical compartment) and incubated for 2 h at 37°C and FITC-dextran levels in the bottom chamber well (basal compartment) was measured. (B) EpiIntestinal 3D microtissues were treated with iAs3+ (1, 5 μM) in presence or absence of UroA (10μM) for 14 days. The supernatants were collected every 3 days for 14 days and pooled. LDH release in these cumulative supernatants at day 14 was measured. (C) IL-8 levels and (D) TNFα levels in supernatants were measured. *p < 0.05, **p < 0.01, ***p < 0.001, Statistics performed using one-way ANOVA in GraphPad Prism software. Error bar, mean ± SEM (n=4).

Article Snippet: Primary human small intestinal epithelial cultures EpiIntestinal 3D microtissues (SMI-100 and SMI-196) were obtained from MatTek Corporation (Ashland, MA, USA) and cultured according to the manufacturer’s instructions (24-well and 96-well format respectively) in a specially formulated culture medium (MatTek Corporation).

Techniques: Incubation, Software

Journal: Mutagenesis

Article Title: Reconstructed human intestinal comet assay, a possible alternative in vitro model for genotoxicity assessment

doi: 10.1093/mutage/gead011

Figure Lengend Snippet: RICom assay treatment timeline of EpiIntestinal™ microtissues. The test chemicals are administered three times: the second administration is 24 h after the first administration. The last administration is 21 h after the second administration (at 3 h before tissue harvesting). Tissues are exposed to test chemicals for 48 h in total.

Article Snippet: Specifically, we adapted a comet assay protocol (RICom) for use with reconstructed 3D EpiIntestinalTM microtissues (MatTek Corporation) and demonstrated that this protocol offers a suitable technology platform for identifying ingested genotoxic hazards for human intestinal tissues.

Techniques:

Journal: Mutagenesis

Article Title: Reconstructed human intestinal comet assay, a possible alternative in vitro model for genotoxicity assessment

doi: 10.1093/mutage/gead011

Figure Lengend Snippet: Historical negative range for solvent control used in RICom assay. Percentage of tail DNA (blue dots) in individual EpiIntestinal™ microtissues treated with solvent control (1% DMSO or 10% H 2 O). Each dot represents data from a single treated tissue. Lower red-dashed line denotes historical mean value. Upper blue-dashed line denotes two standard deviations upper limit. Data are reported as median % tail DNA.

Article Snippet: Specifically, we adapted a comet assay protocol (RICom) for use with reconstructed 3D EpiIntestinalTM microtissues (MatTek Corporation) and demonstrated that this protocol offers a suitable technology platform for identifying ingested genotoxic hazards for human intestinal tissues.

Techniques: Solvent, Control