Human intestinal organoids (HIOs) are a promising in vitro model consisting of different intestinal cell types with a 3D microarchitecture resembling native tissue. In the current study, we aimed to assess the expression of the most common intestinal CYP enzymes in a human induced pluripotent stem cell (hiPSC)-derived HIO model, and the suitability of that model to study chemical-induced changes in CYP expression and activity. We compared this model with the commonly used human colonic adenocarcinoma cell line Caco-2 and with a human primary intestinal epithelial cell (IEC)-based model, closely resembling in vivo tissue. We optimized an existing protocol to differentiate hiPSCs into HIOs and demonstrated that obtained HIOs contain a polarized epithelium with tight junctions consisting of enterocytes, goblet cells, enteroendocrine cells and Paneth cells. We extensively characterized the gene expression of CYPs and activity of CYP3A4/5, indicating relatively high gene expression levels of the most important intestinal CYP enzymes in HIOs compared to the other models. Furthermore, we showed that CYP1A1 and CYP1B1 were induced by β-naphtoflavone in all three models, whereas CYP3A4 was induced by phenobarbital and rifampicin in HIOs, in the IEC-based model (although not statistically significant), but not in Caco-2 cells. Interestingly, CYP2B6 expression was not induced in any of the models by the well-known liver CYP2B6 inducer phenobarbital. In conclusion, our study indicates that hiPSC-based HIOs are a useful in vitro intestinal model to study biotransformation of chemicals in the intestine.
A set
of quality assurance/quality control (QA/QC) criteria for
nontargeted measurement of pesticide exposure markers in a large-scale
study of human urine has been proposed and applied across five laboratories
within the HBM4EU project. Quality control material, including reference
standards and fortified pooled urine samples (QC urine) were prepared
in a centralized way and distributed across participants to monitor
analytical performance and consistency of the liquid chromatography
coupled to high-resolution mass spectrometry data generated with a
harmonized workflow. Signal intensities, mass accuracy, and retention
times of selected QA/QC markers covering a broad range of physicochemical
properties were monitored across QC solvent standards, QC urine samples,
study urine samples, and procedural blanks, setting acceptance thresholds
for repeatability and accuracy. Overall, results showed high repeatability
of the collected data. The RSDs of the signal intensities were typically
below 20–30% in QC and study samples, with good stability of
the chromatographic separation (retention time drift within 2–4
s intrabatch and 5 s interbatch) and excellent mass accuracy (average
error < 2 ppm). The use of the proposed criteria allowed for the
identification of handling errors, instrumental issues, and potential
batch effects. This is the first elaboration of harmonized QA/QC criteria
applied across multiple laboratories to assess the quality of data
generated by nontargeted analysis of human samples.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.