In the context of human airway organoid formation in ALI Medium, do the matrix domes spread on Nunclon Sphera™ plates, and do they remain attached over a 3–4 week culture period?

For the formation of human airway organoids cultured in ALI Medium, proper handling of the matrix is critical to obtain well‑defined, drop‑like domes.
To avoid matrix spreading, the plate should be preheated for at least 2 hours at 37°C, and the gel-cell mixture should be transferred quickly into the wells. Using a warming plate under the laminar flow hood during transfer can further help maintain dome integrity.
In our testing, Nunclon Sphera™ wells were not ideal for this airway organoid system, as the domes did not remain firmly attached. Instead, they were able to move along the bottom of the well and became misshaped over time. Importantly, this altered dome shape did not negatively affect airway organoid growth, and organoids were successfully cultured for up to 4 weeks.
If dome movement is acceptable for the application, extra care should be taken during medium changes. We recommend holding the plate at an angle and gently pipetting to remove spent medium. Vacuum aspiration should be avoided, as it can damage or destroy the dome.