A Fluorescence-based Assay for Characterization and Quantification of Lipid Droplet Formation in Human Intestinal Organoids

作者： Jorik M. van Rijn1,2,3, Marliek van Hoesel1,2, Sabine Middendorp1,2 1Division of Pediatrics, Department of Pediatric Gastroenterology, Wilhelmina Children's Hospital,University Medical Center Utrecht, Utrecht University, 2Regenerative Medicine Center,University Medical Center Utrecht, Utrecht University, 3Science for Life Laboratory, Department of Medical Biochemistry and Microbiology,Uppsala University

简介：

Overview

This protocol describes methods to quantify lipid droplet formation in human intestinal organoids. It serves as a high-throughput screening platform to test drugs that modulate lipid droplet formation.

Key Study Components

Area of Science

• Neuroscience
• Cell Biology
• Pharmacology

Background

• Lipid droplets are important for lipid metabolism.
• Human intestinal organoids provide a physiologically relevant model.
• The assay uses a lipid droplet-specific dye.
• It can be applied to various organoid cultures.

Purpose of Study

• To quantify lipid droplet formation in response to fatty acids.
• To screen for patient-specific drug candidates.
• To understand lipid metabolism in different cell types.

Methods Used

• Utilization of biopsy-derived intestinal organoids.
• Application of a lipid droplet-specific dye.
• High-throughput screening for drug modulation.
• Quantification of lipid droplet formation.

Main Results

• The assay provides stable and accurate results.
• It allows for the identification of drugs affecting lipid droplet formation.
• Can be adapted for other organoid cultures.
• Supports research in lipid metabolism across various conditions.

Conclusions

• The protocol is effective for studying lipid droplet dynamics.
• It has potential applications in drug discovery.
• Provides insights into lipid metabolism in intestinal cells.

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