High Content Imaging Applications of Advanced and Translational Disease Models in Drug Discovery and Development

Royal Society of Chemistry eBooks, Journal Year: 2025, Volume and Issue: unknown, P. 168 - 208

Published: April 30, 2025

There has been increasing interest in disease models with enhanced physiological fidelity. This led to the development of new methods for generating advanced utilizing primary cells and renewable sources, such as induced pluripotent stem organoids. Furthermore, combining these types high content imaging is expected positively impact all stages drug discovery pipeline. Since data rich assays can uncover nuanced cellular response perturbation. In this review, we focus on recent application models, covering general considerations cell source, culture format screening, preclinical studies translational applications, functional precision medicine approaches.

Language: Английский

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Application of new approach methodologies for nonclinical safety assessment of drug candidates

Nature Reviews Drug Discovery, Journal Year: 2025, Volume and Issue: unknown

Published: May 2, 2025

Language: Английский

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Brain organoids engineered to give rise to glia and neural networks after 90 days in culture exhibit human-specific proteoforms

Frontiers in Cellular Neuroscience, Journal Year: 2024, Volume and Issue: 18

Published: May 9, 2024

Human brain organoids are emerging as translationally relevant models for the study of human health and disease. However, it remains to be shown whether human-specific protein processing is conserved in organoids. Herein, we demonstrate that cell fate composition unguided dictated by culture conditions during embryoid body formation, at this stage can optimized result presence glia-associated proteins neural network activity early three-months vitro. Under these conditions, generated from induced pluripotent stem cells (iPSCs) derived male–female siblings similar growth rate, size, total content, exhibit minimal batch-to-batch variability metabolism. A comparison neuronal, microglial, macroglial (astrocyte oligodendrocyte) markers reveals profiles more autopsied cortical cerebellar than those mouse samples, providing first demonstration largely Thus, our organoid protocol provides four major types appear process a manner very brain, they do so half time required other protocols. This unique copy basic characteristics lay foundation future studies aiming investigate brain-specific patterning (e.g., isoforms, splice variants) well modulate glial neuronal processes an situ -like environment.

Language: Английский

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Integrated Molecular-Phenotypic Profiling Reveals Metabolic Control of Morphological Variation in Stembryos

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Dec. 5, 2023

SUMMARY Mammalian stem-cell-based models of embryo development (stembryos) hold great promise in basic and applied research. However, considerable phenotypic variation despite identical culture conditions limits their potential. The biological processes underlying this seemingly stochastic are poorly understood. Here, we investigate the roots by intersecting transcriptomic states morphological history individual stembryos across stages modeling post-implantation early organogenesis. Through machine learning integration time-resolved single-cell RNA-sequencing with imaging-based quantitative profiling, identify features predictive end-state. Leveraging power revealed that imbalance oxidative phosphorylation glycolysis results aberrant morphology a neural lineage bias can be corrected metabolic interventions. Collectively, our work establishes divergent as drivers variation, offers broadly applicable framework to chart predict organoid systems. strategy leveraged control processes, ultimately increasing reproducibility vitro Highlights Time-resolved charting hundreds generates molecular fingerprints Machine identifies end-state Early cellular composition Metabolic interventions tune stembryo correct derailment differentiation outcomes

Language: Английский

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MAGIC matrices: freeform bioprinting materials to support complex and reproducible organoid morphogenesis

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Feb. 5, 2024

Organoids are powerful models of tissue physiology, yet their applications remain limited due to relatively simple morphology and high organoid-to-organoid structural variability. To address these limitations we developed a soft, composite yield-stress extracellular matrix that supports optimal organoid morphogenesis following freeform 3D bioprinting cell slurries at tissue-like densities. The material is designed with two temperature regimes: 4 °C it exhibits reversible behavior support long printing times without compromising viability. When transferred culture 37 °C, the cross-links similar viscoelasticity plasticity basement membrane extracts such as Matrigel. We first characterize rheological properties MAGIC matrices optimize morphogenesis, including low stiffness stress relaxation. Next, combine this custom piezoelectric printhead allows more reproducible robust self-organization from uniform spatially organized "seeds." apply for high-throughput generation intestinal, mammary, vascular, salivary gland, brain arrays structurally those grown in pure Matrigel, but exhibit dramatically improved homogeneity size, shape, maturation time, efficiency morphogenesis. flexibility method enabled fabrication fully microphysiological systems, perfusable tubes experience cyclic strain response pressurization. Furthermore, reproducibility structure increased statistical power drug assay by up 8 orders-of-magnitude given number comparisons. Combined, advances lay foundation efficient complex morphologies canalizing both space time.

Language: Английский

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