I am curious about scalability. For this to work for drug discovery, throughput must be fairly high. I would love to hear your thoughts about how to increase throughput.

Answer

The nature of organoids provides an excellent basis for drug discovery. As referenced in the question, a large volume of organoids would be required to move organoids into a high throughput screening environment. Although there are still currently challenges in this area, including labor-intensive work and time, there are protocols that briefly introduce how organoids can be expanded.

Application Notes and Review Articles:

• A Novel Method for Generating Single, Intestinal Organoids for High Throughput Screening
• Generation of Cerebral Organoids from Human Pluripotent Stem Cells
• A Simple Bioreactor-Based Method to Generate Kidney Organoids from Pluripotent Stem Cells

For scalability, organoids can be plated in single well plates. When preparing organoids for a drug screen, an extra split in which organoids are processed 1:1 and plated again for 1-2 days canincrease the number of organoids significantly. This gives the organoids time to recover from the stress of disruptions, and let them reach the appropriated size for screening activities (20-70 µm). We  have not had good experience plating organoids for drug screening as single cells, since not all single cells are able to grow as organoids. During the manipulation of the organoids for drug screening, we recommend supplementing the washing medium with RhoKi 10 µm to reduce the stress that generates on organoids that are not surrounded by ECM. We do not recommend storing organoids on ice while collecting all material.

Higher throughput for drug screening can be achieved by studying organoid in HTS microplates (e.g. 384-well plates) coated with Corning Matrigel matrix, using the “sandwich” method or embedded method (refer to Corning CLS Guideline for use SPC-356255-G). You can also refer to the article, “Assay Establishment and Validation of a High-Throughput Screening Platform for Three-Dimensional Patient-Derived Colon Cancer Organoid Cultures”, which reports the establishment and validation of a high-throughput screening platform in a 384-well format for 3D organoid culture derived from colon cancer patients.