Microfluidic Co‐Culture Platform to Recapitulate the Maternal–Placental–Embryonic Axis

Boos, Julia Alicia; Misun, Patrick M.; Brunoldi, Giulia; Furer, Lea A; Aengenheister, Leonie; Modena, Mario M.; Rousset, Nassim; Buerki-Thurnherr, Tina; Hierlemann, Andreas

doi:10.1002/adbi.202100609

Cited by 27 publications

(42 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We may use alternative morphology-based assays of gastruloids to complement the TeraTox readout ( Baillie-Benson et al, 2020 ; Moris et al, 2020 ). Alternatively, sophisticated microphysiological systems may better mimic the maternal-placenta-embryo axis and hence, may recapitulate true embryo exposure levels and give insights into active drug metabolism although drugs do not need to cross the placental barrier to cause fetal harm ( Blundell et al, 2016 , 2018 ; Boos et al, 2021 ). In the future, they may replace the 3D EBs in TeraTox .…”

Section: Discussionmentioning

confidence: 99%

Optimization of the TeraTox Assay for Preclinical Teratogenicity Assessment

Jaklin

Zhang

Schäfer

et al. 2022

Toxicological Sciences

View full text Add to dashboard Cite

Current animal-free methods to assess teratogenicity of drugs under development still deliver high numbers of false negatives. To improve the sensitivity of human teratogenicity prediction, we characterized the TeraTox test, a newly developed multi-lineage differentiation assay using 3D human induced pluripotent stem cells. TeraTox produces as primary output concentration-dependent cytotoxicity and altered gene expression induced by each test compound. These data are fed into an interpretable machine-learning model to perform prediction, which relates to the concentration-dependent human teratogenicity potential of drug candidates. We applied TeraTox to profile 33 approved pharmaceuticals and 12 proprietary drug candidates with known in vivo data. Comparing TeraTox predictions with known human or animal toxicity, we report an accuracy of 69% (specificity: 53%, sensitivity: 79%). TeraTox performed better than two quantitative structure-activity relationship models, and had a higher sensitivity than the murine embryonic stem cell test (accuracy: 58%, specificity: 76%, sensitivity: 46%) run in the same laboratory. The overall prediction accuracy could be further improved by combining TeraTox and mEST results. Furthermore, patterns of altered gene expression revealed by TeraTox may help grouping toxicologically similar compounds and possibly deducing common modes of action. The TeraTox assay and the dataset described here therefore represent a new tool and a valuable resource for drug teratogenicity assessment.

show abstract

Section: Discussionmentioning

confidence: 99%

Optimization of the TeraTox Assay for Preclinical Teratogenicity Assessment

Jaklin

Zhang

Schäfer

et al. 2022

Toxicological Sciences

View full text Add to dashboard Cite

show abstract

“…Therefore, such studies can be performed without the need for prior isolation of biomolecules, similar to the utilization of the original chip design [ 25 ]. Other microfluidic technologies that have been developed in recent years to study cell communication between co-cultured populations rely on more complicated fabrication methods and the use of non-physiological barriers, such as filters or channels with reduced width, and also do not enable cell migration studies [ 43 , 44 , 45 ].…”

Section: Discussionmentioning

confidence: 99%

A Microfluidic Platform to Monitor Real-Time Effects of Extracellular Vesicle Exchange between Co-Cultured Cells across Selectively Permeable Barriers

Mason

Bush

Agrawal

et al. 2022

IJMS

View full text Add to dashboard Cite

Exosomes and other extracellular vesicles (EVs) play a significant yet poorly understood role in cell–cell communication during homeostasis and various pathological conditions. Conventional in vitro and in vivo approaches for studying exosome/EV function depend on time-consuming and expensive vesicle purification methods to obtain sufficient vesicle populations. Moreover, the existence of various EV subtypes with distinct functional characteristics and submicron size makes their analysis challenging. To help address these challenges, we present here a unique chip-based approach for real-time monitoring of cellular EV exchange between physically separated cell populations. The extracellular matrix (ECM)-mimicking Matrigel is used to physically separate cell populations confined within microchannels, and mimics tissue environments to enable direct study of exosome/EV function. The submicron effective pore size of the Matrigel allows for the selective diffusion of only exosomes and other smaller EVs, in addition to soluble factors, between co-cultured cell populations. Furthermore, the use of PEGDA hydrogel with a very small pore size of 1.2 nm in lieu of Matrigel allows us to block EV migration and, therefore, differentiate EV effects from effects that may be mediated by soluble factors. This versatile platform bridges purely in vitro and in vivo assays by enabling studies of EV-mediated cellular crosstalk under physiologically relevant conditions, enabling future exosome/EV investigations across multiple disciplines through real-time monitoring of vesicle exchange.

show abstract

“…Random forest, which is an ensemble learning method, is better at detecting such heterogeneous signals than linear regression. to cross the placental barrier to cause fetal harm (Blundell et al 2016;Blundell et al 2018;Boos et al 2021). In the future, they may replace the 3D embryoid bodies in TeraTox.…”

Section: Discussionmentioning

confidence: 99%

“…We may use alternative morphology-based assays of gastruloids to complement the TeraTox readout (Baillie-Benson et al 2020; Moris et al 2020). Alternatively, sophisticated micro-physiological systems may better mimic the maternal-placenta-embryo axis and hence, may recapitulate true embryo exposure levels and give insights into active drug metabolism although drugs do not need to cross the placental barrier to cause fetal harm (Blundell et al 2016; Blundell et al 2018; Boos et al 2021). In the future, they may replace the 3D embryoid bodies in TeraTox .…”

Section: Discussionmentioning

confidence: 99%

Optimization of the TeraTox assay for preclinical teratogenicity assessment

Jaklin

Zhang

Schaefer

et al. 2021

Preprint

View full text Add to dashboard Cite

Teratogenicity poses severe threats to patient safety. Stem-cell-based in vitro systems are promising tools to predict human teratogenicity. However, current in vitro assays are limited because they either capture effects on a certain germ layer, or focus on a subset of predictive markers. Here we report the characterization and critical assessment of TeraTox, a newly developed multi-lineage differentiation assay using 3D human induced pluripotent stem cells. TeraTox probes stem-cell derived embryoid bodies with two endpoints, one quantifying cytotoxicity and the other inferring the teratogenicity potential with gene expression as a molecular phenotypic readout. To derive teratogenicity potentials from gene expression profiles, we applied both unsupervised machine-learning tools including factor analysis and supervised tools including classification and regression. To identify the best predictive model for the teratogenicity potential that is explainable, we systematically tested 64 machine-learning model architectures and identified the optimal model, which uses expression of 77 representative germ-layer genes, summarized by 10 latent germ-layer factors, as input for random-forest regression. We combined measured cytotoxicity and inferred teratogenicity potential to predict concentration-dependent teratogenicity profiles of 33 approved pharmaceuticals and 12 proprietary drug candidates with known in vivo data. Compared with the mouse embryonic stem cell test, which has been in routine use for more than a decade, the TeraTox assay shows higher sensitivity, particularly towards teratogens impairing ectodermal development or stem-cell renewal, and a more balanced prediction performance. We envision that further refinement and development of TeraTox has the potential to reduce and replace animal research in drug discovery and to improve preclinical assessment of teratogenicity.

show abstract

Microfluidic Co‐Culture Platform to Recapitulate the Maternal–Placental–Embryonic Axis

Cited by 27 publications

References 43 publications

Optimization of the TeraTox Assay for Preclinical Teratogenicity Assessment

Optimization of the TeraTox Assay for Preclinical Teratogenicity Assessment

A Microfluidic Platform to Monitor Real-Time Effects of Extracellular Vesicle Exchange between Co-Cultured Cells across Selectively Permeable Barriers

Optimization of the TeraTox assay for preclinical teratogenicity assessment

Contact Info

Product

Resources

About