Journal: Toxics

Article Title: Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models

doi: 10.3390/toxics10100566

Figure Lengend Snippet: Experimental design. Schematic diagram describing the chemical mixture tested across different in vitro liver models (suspension, 2D sandwich, and OrganoPlate ® 2-lane 96) and types of chemical analyses used in this study.

Article Snippet: For evaluation of drug metabolism, iHep co-cultured with THP-1/HMEC in the OrganoPlate ® 2-lane 96 were exposed on days 8 and 12 of culture to either 1 or 5 µM mixture of 20 pesticides.

Techniques: In Vitro

Journal: Toxics

Article Title: Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models

doi: 10.3390/toxics10100566

Figure Lengend Snippet: Albumin and lactate dehydrogenase (LDH) in traditional 2D sandwich culture ( A ) and OrganoPlate ® 2-lane 96 ( B ). Data are plotted as box (interquartile range) and whiskers (min–max range) with individual data points shown; horizontal line is median. One asterisk (*) denotes statistical differences at p < 0.05 (one-way ANOVA with Dunnett’s multiple comparisons test). Two asterisks denote differences at p < 0.01, and three asterisks denote statistical differences at p < 0.001.

Article Snippet: For evaluation of drug metabolism, iHep co-cultured with THP-1/HMEC in the OrganoPlate ® 2-lane 96 were exposed on days 8 and 12 of culture to either 1 or 5 µM mixture of 20 pesticides.

Techniques:

Journal: Toxics

Article Title: Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models

doi: 10.3390/toxics10100566

Figure Lengend Snippet: ( A ) In vitro hepatocyte clearance of chemicals in suspension culture (squares), sandwich culture (circles), OrganoPlate ® 2-lane 96 (triangles), and httk (diamonds). Plotted are in vitro hepatocyte clearances (mean) of each chemical listed (the number in brackets indicates the LogP of each compound). Experiments were conducted using induced pluripotent stem cell-derived hepatocytes (iHep). Each chemical was tested at 1 µM in a mixture setting. Clearance values reported for each chemical in httk R package version 1.10.1 were tested using primary human hepatocyte suspensions at 10 µM in a single chemical setting. ( B ) 3D plot showing hepatocyte clearance of each chemical across the three in vitro models used in this study. Axes represent in vitro clearance values for each compound in a 1 µM mixture. All data points for each model can be found in .

Article Snippet: For evaluation of drug metabolism, iHep co-cultured with THP-1/HMEC in the OrganoPlate ® 2-lane 96 were exposed on days 8 and 12 of culture to either 1 or 5 µM mixture of 20 pesticides.

Techniques: In Vitro, Derivative Assay

Journal: Toxics

Article Title: Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models

doi: 10.3390/toxics10100566

Figure Lengend Snippet: Experimental design. Schematic diagram describing the chemical mixture tested across different in vitro liver models (suspension, 2D sandwich, and OrganoPlate ® 2-lane 96) and types of chemical analyses used in this study.

Article Snippet: We used iHep suspensions and 2D sandwich cultures, and a microphysiological system OrganoPlate ® 2-lane 96 (Mimetas TM ) that also included endothelial cells and THP-1 cell-derived macrophages.

Techniques: In Vitro

Journal: Toxics

Article Title: Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models

doi: 10.3390/toxics10100566

Figure Lengend Snippet: Albumin and lactate dehydrogenase (LDH) in traditional 2D sandwich culture ( A ) and OrganoPlate ® 2-lane 96 ( B ). Data are plotted as box (interquartile range) and whiskers (min–max range) with individual data points shown; horizontal line is median. One asterisk (*) denotes statistical differences at p < 0.05 (one-way ANOVA with Dunnett’s multiple comparisons test). Two asterisks denote differences at p < 0.01, and three asterisks denote statistical differences at p < 0.001.

Article Snippet: We used iHep suspensions and 2D sandwich cultures, and a microphysiological system OrganoPlate ® 2-lane 96 (Mimetas TM ) that also included endothelial cells and THP-1 cell-derived macrophages.

Techniques:

Journal: Toxics

Article Title: Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models

doi: 10.3390/toxics10100566

Figure Lengend Snippet: ( A ) In vitro hepatocyte clearance of chemicals in suspension culture (squares), sandwich culture (circles), OrganoPlate ® 2-lane 96 (triangles), and httk (diamonds). Plotted are in vitro hepatocyte clearances (mean) of each chemical listed (the number in brackets indicates the LogP of each compound). Experiments were conducted using induced pluripotent stem cell-derived hepatocytes (iHep). Each chemical was tested at 1 µM in a mixture setting. Clearance values reported for each chemical in httk R package version 1.10.1 were tested using primary human hepatocyte suspensions at 10 µM in a single chemical setting. ( B ) 3D plot showing hepatocyte clearance of each chemical across the three in vitro models used in this study. Axes represent in vitro clearance values for each compound in a 1 µM mixture. All data points for each model can be found in .

Article Snippet: We used iHep suspensions and 2D sandwich cultures, and a microphysiological system OrganoPlate ® 2-lane 96 (Mimetas TM ) that also included endothelial cells and THP-1 cell-derived macrophages.

Techniques: In Vitro, Derivative Assay

Journal: Bioengineering

Article Title: Reproducibility and Robustness of a Liver Microphysiological System PhysioMimix LC12 under Varying Culture Conditions and Cell Type Combinations

doi: 10.3390/bioengineering10101195

Figure Lengend Snippet: General study designs for the experiments performed. The range of experimental conditions that was utilized in this study to evaluate PhysioMimix LC-12 and other liver in vitro models. See detailed experimental protocols for each experiment detailed in . Day 0 corresponded to the day when the cells were seeded into devices and media movement commenced (for microfluidic models) or cells were seeded into multi-well plates (for static cultures). In the PhysioMimix LC-12, media flow through the device/scaffolds without cells was initiated before cells were seeded, according to the manufacturer’s protocols. Media changes with or without CYP3A4 assay and the timing of drug/chemical additions are indicated (see symbols legend in the inset). ( A ) The most typical PhysioMimix LC-12 study design spanning a total of 14 days in culture. In the experiments that used NPCs, they were started in culture 2 days prior to the seeding of all cells into the device and initiation of flow. The asterisks (*, **) denote that different types of NPCs were used for PHHs and iHeps 2.0 (see ). ( B ) The PhysioMimix LC-12 study extended to 28 days. ( C ) Studies with iHeps 2.0 in the PhysioMimix LC-12 model. ( D ) Static culture in 384-well plates for iHeps 2.0. ( E ) Static cultures in 96-well plates for PHHs (HU8373 donor only). ( F ) Monoculture PHH (HU8373) study in a LAMPS model. ( G ) Study with iHeps 2.0 in the OrganoPlate 2-lane 96 (Mimetas, Leiden, The Netherlands) model. ( H ) Study of Trovafloxacin +/− LPS in PhysioMimix LC-12 model. ( I ) Study of the metabolism of a pesticide mixture in PhysioMimix LC-12 model.

Article Snippet: For almost all examined compounds , the disappearance of the parent molecules was far more evident in the experiments with HU8373 PHH in PhysioMimix LC12 when compared to iHeps 2.0 cultured in various models (suspension, 2D culture or OrganoPlate ® 2-lane 96).

Techniques: In Vitro

Journal: Bioengineering

Article Title: Reproducibility and Robustness of a Liver Microphysiological System PhysioMimix LC12 under Varying Culture Conditions and Cell Type Combinations

doi: 10.3390/bioengineering10101195

Figure Lengend Snippet: Comparison of basal function across different cell sources and models. ( A ) Albumin and urea secretion, CYP3A4 activity, and LDH leakage in the PhysioMimix LC12 under monoculture conditions across 5 different PHH donors and iPSC-derived hepatocytes (iHeps 2.0). ( B ) Albumin and urea secretion, CYP3A4 activity, and LDH leakage in 2D culture or other liver MPS models (LAMPS or OrganoPlate ® 2-lane 96) under monoculture conditions using either PHH donor HU8373 or iHeps. ( C ) Albumin secretion and CYP3A4 activity in the PhysioMimix LC12 under monoculture and co-culture conditions using PHH donor HU8373 over 28 days of culture. The number of experiments included in each condition of use with a range of individual chips in each experiment is indicated in the labels.

Article Snippet: For almost all examined compounds , the disappearance of the parent molecules was far more evident in the experiments with HU8373 PHH in PhysioMimix LC12 when compared to iHeps 2.0 cultured in various models (suspension, 2D culture or OrganoPlate ® 2-lane 96).

Techniques: Comparison, Activity Assay, Derivative Assay, Co-Culture Assay

Journal: Bioengineering

Article Title: Reproducibility and Robustness of a Liver Microphysiological System PhysioMimix LC12 under Varying Culture Conditions and Cell Type Combinations

doi: 10.3390/bioengineering10101195

Figure Lengend Snippet: Metabolism of a mixture of 20 pesticides across different cell sources and models. Metabolism is shown as % parent compound remaining after 48 h in the PhysioMimix LC-12 using PHH (donor HU8373) + THP-1 with either 1 µM (green squares) or 5 µM (green diamonds) equimolar mixture and compared to the published results in of experiments with iPSC-derived hepatocyte (iHeps 2.0) suspension, 2D culture, and OrganoPlate ® 2-lane 96 (abbreviated as “Mimetas”) experiments. Data shown are mean ± standard deviation (n = 3 in each condition).

Article Snippet: For almost all examined compounds , the disappearance of the parent molecules was far more evident in the experiments with HU8373 PHH in PhysioMimix LC12 when compared to iHeps 2.0 cultured in various models (suspension, 2D culture or OrganoPlate ® 2-lane 96).

Techniques: Pesticides, Derivative Assay, Suspension, Standard Deviation