I'm growing mammospheres according to this protocol:

- Media 0% serum (DMEM, or RPMI according to the cell line)

- B27 1% or 1X

- Insulin 5 ug/mL

- bFGF 20 ng/mL

- Hydrocrotisone 0.5 ug/mL

Does anyone know another recipe for mammospheres/tumorospheres which could help me out?

Besides I ran out of B27, any other factor that can be used for its replacement? As far as I know and experienced, cells aren't being attached to themselves neither forming spheroids without this B27.

Thank you for your help,

Santiago

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