We are using the media from Hans Clever’s paper to grow hepatocyte-based organoids. How often do you recommend media changes? We are seeing that some of our organoids are dying right after media changes, every 2-3 days.

Answer

To support organoid growth, we recommend refreshing media every 2-3 days. Some of the compounds used are not very stable after several days at 37°C. The observation that organoids die right after medium refreshment suggests that a revision or optimization to the media formulation is needed. You also may need to optimize the final concentration of reagents, since the death of the organoids is unexpected if these formulations are all correct.