Analysis of Reproducibility and Robustness of a Renal Proximal Tubule Microphysiological System OrganoPlate® 3-lane 40 for In Vitro Studies of Pharmacokinetics and Toxicodynamics
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PROVIDER: PRJNA976205 | ENA |
REPOSITORIES: ENA
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Project description:Microphysiological systems are a promising new experimental tool for drug and chemical safety evaluation. These models are rapidly evolving, and concerns exist about the reproducibility, robustness and relative benefits when compared to traditional in vitro cultures. This study used OrganoPlate® 3-lane 40, a microfluidics-based model of the renal proximal tubule and compared it to multi-well static cultures using a number of human renal proximal tubule epithelial cell (RPTEC) types. We followed previously published protocols but adapted them to the varying cell proliferation rates of different RPTECs; primary cells required up to 7 days in culture before experiments, while some immortalized cell lines could be used in 3 days. Function of various tubule-specific transporters, permeability of small molecules (cisplatin, tenofovir and perfluorooctanoic acid) and fluorescent dextrans (60-150 kDa), basal and chemical-effected gene expression, and cell viability were tested. Different RPTEC types and culture conditions (OrganoPlate® 3-lane 40 vs multi-well plates) were compared. We found that there are advantages offered by OrganoPlate® 3-lane 40 as compared to multi-well cultures – presence of media flow, albeit intermittent, and fairly high throughput. However, this model appears to offer only limited (e.g., MRP-mediated transport) advantages in terms of either gene expression or transporter function when compared to the multi-well plate culture conditions. It can also be used to study cellular uptake and direct toxic effects of small molecules; still, it may have limited utility for studies of pharmaco-/toxico-kinetics because drug transport between blood and tubule compartments is considerably affected by the gel barrier.
2023-11-08 | GSE233406 | GEO
Project description:Microphysiological systems are an emerging area of in vitro drug development, and their independent evaluation is important for wide adoption and use. The primary goal of this study was to test reproducibility and robustness of a renal proximal tubule microphysiological system, OrganoPlate 3-lane 40, as an in vitro model for drug transport and toxicity studies. This microfluidic model was compared with static multiwell cultures and tested using several human renal proximal tubule epithelial cell (RPTEC) types. The model was characterized in terms of the functional transport for various tubule-specific proteins, epithelial permeability of small molecules (cisplatin, tenofovir, and perfluorooctanoic acid) versus large molecules (fluorescent dextrans, 60-150 kDa), and gene expression response to a nephrotoxic xenobiotic. The advantages offered by OrganoPlate 3-lane 40 as compared with multiwell cultures are the presence of media flow, albeit intermittent, and increased throughput compared with other microfluidic models. However, OrganoPlate 3-lane 40 model appeared to offer only limited (eg, MRP-mediated transport) advantages in terms of either gene expression or functional transport when compared with the multiwell plate culture conditions. Although OrganoPlate 3-lane 40 can be used to study cellular uptake and direct toxic effects of small molecules, it may have limited utility for drug transport studies. Overall, this study offers refined experimental protocols and comprehensive comparative data on the function of RPETCs in traditional multiwell culture and microfluidic OrganoPlate 3-lane 40, information that will be invaluable for the prospective end-users of in vitro models of the human proximal tubule.
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Project description:Current research on metabolic disorders and diabetes relies on animal models because multi-organ diseases cannot be well studied with the standard in vitro assays. Here, we connect models of key metabolic organs, pancreas and liver, on a microfluidic chip to enable diabetes research in a human-based in vitro system. Aided by mechanistic mathematical modelling, we show that hyperglycemia and high cortisone induce glucose dysregulation in the pancreas-liver microphysiological system (MPS) mimicking a diabetic phenotype seen in patients with glucocorticoid-induced diabetes. In this diseased condition, pancreas-liver MPS displays beta-cell dysfunction, steatosis, elevated ketone-body secretion, increased glycogen storage, and upregulated gluconeogenic gene expression. In turn, a physiological culture condition maintains the glucose tolerance and beta cell function. This method was evaluated for reproducibility in two laboratories and was effective in multiple pancreatic islet donors. The model also provides a platform to identify new therapeutic proteins as demonstrated with a combined transcriptome and proteome analysis.
2024-08-10 | PXD052854 | Pride
Project description:Fluorosurfactants are the key components in aqueous film forming foams (AFFF). They provide these fire fighting agents with the required low surface tension and they enable film formation on top of lighter fuels to prevent burn back. Development of effective and environmentally acceptable PFOS alternatives is one of the most important priorities in the fire fighting foam industry. DuPontTM offers the fluorosurfactant mixtures Forafac®1157 and Forafac®1157N for the formulation of AFFFs which are alternatives to the persistent and toxic perfluorooctane sulphonate (PFOS). Ecotoxicological testing of these inadequately documented mixtures is necessary to include them in AFFF hazard and risk assessment. Juvenile turbot (Scophthalmus maximus) was exposed for 14 days to 0.5 and 1.5 mg/l of the fluorosurfactant mixtures used in Forafac®1157 and Forafac®1157N. In a first transcriptomics experiment, microarray analysis revealed differentially expressed gene transcripts which were mainly involved in digestion and in the immune system. This discovery-driven screening approach offered the basis for new hypotheses that were tested in two subsequent experiments in which food intake, energy reserves, growth and a set of haematological parameters were examined. Additionally, effects of the two mixtures were compared to those of PFOS. Based on the results of this study, the mode of action of Forafac®1157N was the activation of the acute phase reaction resulting in increased leukocyte concentrations and the inhibition of growth due to the high energetic cost of toxicant exposure. For Forafac®1157, evidences of immunosuppression were found on the transcriptional level and the altered differential leukocyte profiles indicated that stress was induced in these fish. However, food intake, energy reserves and growth were not compromised, even at high exposure concentrations, which was in contrast to the effects seen after PFOS exposure. Taking into account that Forafac®1157 appeared to be less toxic than PFOS, this mixture could be considered as a more environmentally acceptable PFOS alternative for the use in AFFFs.
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