Incorporating EdU Nucleoside into Organoids: Improved Understanding of Cellular Processes

The incorporation of EdU nucleoside into organoids represents a powerful tool for advancing our understanding of cellular dynamics. Its applications?to study cellular processes such as disease modeling, drug discovery and developmental studies, highlight its versatility and importance in modern biomedical research.? What is EdU? 5-Ethynyl-2’-deoxyuridine (EdU) is a thymidine analog that gets incorporated into DNA during active DNA synthesis. Unlike traditional thymidine analogs like BrdU (Bromodeoxyuridine), EdU offers several advantages due to its ease of detection and minimal disruption to cellular processes. Incorporation of EdU into Organoids EdU incorporation into organoids?[1,2] is a relatively straightforward process: Cultivation: Organoids are grown under standard conditions until they reach the desired developmental stage.

Labeling: EdU is added to the culture medium, where it is taken up by proliferating cells and incorporated into their DNA during replication.

Detection: Using click chemistry reaction, EdU can be detected with a fluorescent azide, enabling researchers to visualize and quantify cell proliferation with high precision. ?

Function and Benefits of EdU Incorporation

• Cell Proliferation Studies: EdU allows for precise measurement of cell proliferation rates within organoids. This is crucial for understanding growth patterns, tissue regeneration, and responses to various treatments.
• ?Minimal Toxicity: EdU is less toxic compared to other nucleotide analogs, ensuring that the integrity and function of the organoid are maintained during experiments.
• High Sensitivity: The click chemistry reaction used to detect EdU incorporation is highly sensitive and specific, providing clear and reliable results.

Applications of EdU in Organoid Research

• Cancer Research: By incorporating EdU, researchers can study tumor growth dynamics and the effects of anti-cancer drugs on cell proliferation within tumor organoids. This can lead to the identification of more effective treatment regimens.
• Developmental Biology: EdU helps in tracking cell division and differentiation during organoid development, shedding light on the fundamental processes of organogenesis.
• Toxicology: EdU incorporation allows for the assessment of toxic effects of various compounds on cell proliferation, aiding in the safety evaluation of new drugs and chemicals.
• Regenerative Medicine: Understanding cell proliferation within organoids is essential for developing regenerative therapies. EdU helps in optimizing conditions for tissue regeneration and repair.

Case Study: EdU in Intestinal Organoids

A recent study [1] utilized EdU to investigate cell proliferation in intestinal organoids derived from patient biopsies. The findings revealed distinct proliferation zones corresponding to crypt and villus regions, mirroring in vivo conditions. This breakthrough underscores the potential of EdU incorporation in replicating and studying complex tissue structures and functions.

baseclick’s ClickTech EdU Cell Proliferation Kit To support diverse analytic methods, baseclick offers the ClickTech in vivo EdU Cell Proliferation Kits. These comprehensive kits are designed for various applications, including:

• Imaging: Facilitates the visualization of cell proliferation through fluorescence microscopy.
• Flow Cytometry: Allows for the quantification of proliferating cells in a population.
• High-Throughput Screening: Enables large-scale screening of compounds affecting cell proliferation.

Further Reading:

[1] Li HS et al. Quantification of Proliferative and Dead Cells in Enteroids. J Vis Exp. (2020) ;(155):10.3791/60501. doi: 10.3791/60501. PMID: 32065148; PMCID: PMC10335857. ?

[2] Chnasu Lee et al.? Epithelial Regeneration Ability of Crohn’s Disease Assessed Using Patient-Derived Intestinal Organoids Int. J. Mol. Sci. (2021), 22(11), 6013; https://doi.org/10.3390/ijms22116013

[3] Zhao, Z., et al. Organoids. Nat Rev Methods Primers 2, 94 (2022). https://doi.org/10.1038/s43586-022-00174-y