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