Abstract: Human organoid 3D cultures are models representing a precious asset for biomedical research, from fundamental discovery to precision medicine. Beyond access to the primary tissue and line establishment, the main limitations to the use of human-derived organoid models are the amount of time and skills required to maintain these cultures and also the costs associated with their production/maintenance (staff time and reagents). We describe a successful way to increase our organoid culture capacities while significantly decreasing the associated costs using a bioreactor. We measured the growth rate (i.e cell counting), compared the morphology (i.e H&E staining) and quantified the culture costs (staff time hours and reagents) in the bioreactor against our standard culture method (100% Matrigel embedded organoids overlayed with complete media in 24 well-plates). We tested different conditions in the bioreactor (various inoculation cell numbers and addition of 0%, 2% or 4% Matrigel) and compared with matching plates (standard culture condition). Our results generated using patient-derived colon and breast cancer organoids demonstrated that producing 10 million of single cell organoids was 4 times faster and 5 times cheaper in the bioreactor than in standard culture. Other non-quantitative metrics include the ease of use, the flexibility of the system allowing to successfully culture organoids from 6ml to 30ml volume of media and starting with a number of single cells as low as 150K. This is particularly valuable when the access to the primary tissue and/or the established organoid line is very limited.
