WEBINAR

Working with Complex 3D Cell Models

Introduction

Part 3 of A Webinar Series: Towards Physiological Relevance - High Content Analysis with 3D Organoids and Spheroids

In this webinar, hear from two leading scientists about their approaches to working with complex 3D models using high-content analysis technologies.

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Optimizing High Content-Screening for Organoid-Derived Microfluidic Intestine Chips

Marianne Kanellias

Emulate Inc, Boston, MA, United States

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High Throughput Screening for Cell-Type-Specific Toxicity in Human Midbrain Organoids

Dr. Jan Bruder

Max Planck Institute for Molecular Biomedicine, Münster, Germany

Abstracts:

Optimizing High Content-Screening for Organoid-Derived Microfluidic Intestine Chips

For imaging Organ-Chip models supporting 3D cellular structures, HCS systems can help increase the throughput while maintaining confocal z-resolution. Discover how Emulate have utilized the Plate Layout and PreciScan capabilities of the Opera Phenix® system to capture the microfluidic channels of primary organoid-seeded Colon and Duodenum Chips for an array of applications.

You will learn about:

  • How to adapt a Plate Layout to Emulate Organ-Chips
  • Utilizing PreciScan to align to the Organ-Chip epithelial channel
  • Optimal acquisition settings for capturing 3D epithelium of Colon and Duodenum-Chips

High Throughput Screening for Cell-Type-Specific Toxicity in Human Midbrain Organoids

Toxicity testing is a critical step in the development and approval of chemical compounds for human contact and consumption. However, existing model systems are often inadequate in predicting human toxicity in vivo because they may not adequately recapitulate human physiology. The complexity of three-dimensional (3D) human organ-like cell culture systems ("organoids") can potentially generate more relevant models of human physiology and disease, including toxicity predictions.

To date, however, the inherent biological heterogeneity and cumbersome generation and analysis of organoids have made efficient, unbiased, high-throughput evaluation of toxic effects in these systems difficult. Recent advances in both standardization and quantitative fluorescence imaging have allowed us to study the toxicity of compound exposure to separate cellular subpopulations in human organoids at the single-cell level in a framework compatible with high-throughput approaches. Screening a library of 84 compounds in standardized human automated midbrain organoids (AMOs) generated from two independent cell lines correctly identified known nigrostriatal toxicities. This approach further identified for the first time the flame retardant 3,3′,5,5′-tetrabromobisphenol A (TBBPA) as a selective toxin for dopaminergic neurons in the context of human midbrain-like tissues.

The results were verified with high reproducibility in more detailed dose-response experiments. Furthermore, we demonstrate higher sensitivity in 3D AMOs than in 2D cultures to the known neurotoxic effects of the pesticide lindane. Overall, the automated nature of our workflow is freely scalable and demonstrates the feasibility of quantitatively assessing cell type-specific toxicity in human organoids in vitro.