Hi,
As harvesting spheroid from primary cells takes longer than cell line and as hanging drop method is best method among others is best for spheroid formation, the changing medium would be an serious issue. if someone used hanging drop for producing spheroids, can someone explain me how and after how many days the medium was changed for their spheroids?
Do we still use such methods? It is too old method. Nowadays, you just have to use ultra low attachment flasks or nunclon sphera flaks and you can easily get sphere.
I would say that it is almost impossible for a media change in hanging drop assay. However, you can try to minimize evaporation by putting water at the bottom of the plate, or making sure the water reservoir in the incubator is properly filled. You can also increase the initial number of cells used for spheroid formation, more cells will ease formation of spheroid better. Try also supplementing your hanging drop media with 0.24% methylcellulose. Good luck.
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