This article explores automating a 3D intestinal organoids culture using an automated cell culture system.
Sponsored Content by Molecular Devices UK LtdReviewed by Aimee MolineuxAug 5 2024 Attrition in the therapeutic pipeline can often be associated with the lack of translational efficacy from the pre-clinical phase to the clinic. Organoids demonstrate the significant potential to change the game in disease modeling and drug screening since they closely resemble tissue structure and functionality and demonstrate more predictive responses to drugs.
Subsequently, the organoids were able to self-organize and develop complex crypt structures. To determine organoid number, size , and optical density, organoids were observed in transmitted light and evaluated using machine learning-based image analysis. For endpoint assay , organoids were stained for viability markers and checked for concentration and time-dependent effects of compounds on healthy intestinal organoids , or patient-derived colorectal cancer organoids .
Cell culture protocols Molecular devices UH has developed a protocol for 3D intestinal organoids* that were taken from primary mouse intestinal cells using existing methods . Cells were cultured and differentiated in line with the STEMCELL Technologies protocol. IntestiCult™ Organoid Growth Medium was applied for cell culture.
Organoid plating was initiated from the organoids suspended in Matrigel which was set into a pre-chilled 96-well deep well block. The suspension was pipetted using four pipette tips and dispensed into a 24-well plate, four tips at a time. Plating into a 96-well format was also tested. For the passaging process, the removal of the media was completed, and Matrigel domes were incubated using Gentle cell dissociation reagent, then the Matrigel domes were broken using rigorous pipetting. The mix was harvested into a 96-deep-well block. Then optional pooling of two wells into one was carried out, followed by centrifugation on an external centrifuge with a speed of 400 g for five minutes.
Results In this study, continuous culture of intestinal organoids was carried out for more than a month. Automated seeding of organoid domes enabled consistent size and precise centering of domes in 24-well plates, which made imaging and image analysis easier and more accurate. Organoids grow as expected throughout the automated culture process, creating protrusions in keeping with typical intestinal organoids morphology.
of organoid analysis: organoid count and skewness over time. Skewness includes a combination of optical density, granularity, and other optical However, this study demonstrates how researchers can prevent certain bottlenecks that come with labor-intensive, complex protocols to increase productivity for drug screening.
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