Organoid Viability Assay

An organoid viability assay is a method for determining whether cells within a three-dimensional organoid remain alive and functional under defined culture conditions. It typically uses indicators of cell membrane integrity, metabolic activity, or both to distinguish viable cells from damaged or dead cells, often through fluorescence or colorimetric measurements. In developmental biology, viability assays help evaluate organoid formation, growth, differentiation, and responses to genetic or environmental changes. They also support optimization of culture conditions and comparison of developmental models by revealing whether observed structural changes reflect normal maturation, cellular stress, or loss of viability.

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JoVE EoE - Assay Techniques

ATP-Based Luciferase Viability Assay: A Homogenous Method to Evaluate the Growth-Inhibitory Potential of Test Agents on Tumor Organoids

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2025

In this video, we demonstrate a rapid, single-addition assay to assess cell viability in patient-derived organoids upon treatment with a test agent using intracellular ATP levels as a marker for cellular metabolic activity. The intracellular ATP levels are measured using the luciferase/luciferin reaction, where the emitted light intensity indicates the cellular viability and, in turn, the test agent's growth-inhibitory potency.

Viability Assays for Cells in Culture

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Cited by 39 •

2014

Therapeutic compounds are often first examined in vitro with viability assays. Blind cell counts by a human observer can be highly sensitive to small changes in cell number but do not assess function. Computerized viability assays, as described here, can assess both structure and function in an objective manner.

XTT Assay: A Colorimetric Assay to Assess Cell Viability and Mitochondrial Activity

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2023

This article describes the protocol for XTT assay which is a colorimetric assay to assess cell viability and mitochondrial activity.

Forskolin-induced Swelling in Intestinal Organoids: An In Vitro Assay for Assessing Drug Response in Cystic Fibrosis Patients

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Cited by 121 •

2017

This protocol describes an assay for measuring CFTR function and CFTR modulator responses in cultured tissue from subjects with cystic fibrosis (CF). Biopsy-derived intestinal organoids swell in a cAMP-driven fashion, a response that is defective (or strongly reduced) in CF organoids and can be restored by exposure to CFTR modulators.

Agarose Spin Down Assay: A Technique to Embed Organoids for Histological Analysis

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2023

In this video, we perform a spin-down assay to embed 3D prostate cancer organoids in agarose. These embedded organoids can be used for further histopathological analysis to study cancer growth and development.

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