Dear all,

I am working with spheroids of U251 and U87 glioblastoma cells. Now, I would like to dissociate the spheroids for single cell flow cytometry analysis. However, either the spheroids are not dissociated, or the single cells are not viable.

Does anyone have a protocol?

Here are some more details

- cells: U87, U251

- spheroids: 4000 cells seeded in suspension with 0.24% methocell, 3-5 days incubation ( +/- 400-600 µm)

- accutase and tryple have been independently used for dissociation; up to 30-60 minutes; tryple also using a rocking shaker; I avoid using trypsin for flow cytometry analyses

- mechanical forces by pipetting up-and-down

Thanks & Best,

Jorrit

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