since spheroid is a microtissue structure, you might also want to try collagenase which we regularly use to digest tumor tissue to get single cells suspension.

As a reference, you can refer to this paper where they use various kinds of collagenase to digest lung tumor tissue

Article Technical Advance: An optimized disaggregation method for hu...

Thanks for your answer, Win.

I found that combination of trypsin and cell dissociate buffer works too.

Hi,Yuan,I'm also trying to disassocaite spheroids by 0.25% tripsin but it didn't work well after long-term culture maybe because the spheroids become more compact , could you tell me how you did this by trypsin and cell dissociate buffer,like the ratio of this two type of reagents,thank you so much!

Hi Yuan Lyu , Using accutase you can get single cell suspension easily from spheroid. This product is available from many sources. Try it and get back

Thanks