I am trying to do a wound healing assay with Schwann cells, but my cells die when I starve them in serum free medium. I have also tried different amounts of FBS, but they seem to die unless there is 5% or higher FBS in the medium. I can decrease FBS to 1% if I keep forskolin and neuregulin in the medium.
Does anyone have experience with Schwann cell wound healing assays and an optimal starvation medium for the assay ? Is it a problem to keep forskolin and neuregulin in the medium during starvation and the assay?
I want to examine migration in NT3 stimulated wt and KO Schwann cells.
All comments or suggestions for improvements are greatly appreciate.
Thanks!
Thank you @Victor Carriel. It is primary Schwann cells I prepare from postnatal rats (P1-3). I typically use the Schwann cells for experiments when they are at passage 3-4.
Which medium are you using?
0.05% FBS should be ok without nrg and forskolin.
Thank you, Jose A. Gomez-Sanchez .
When the cells are thawed, they proliferate in DMEM + 10% FBS + neuregulin and forskolin. Then, when they have formed a confluent monolayer, I wash twice with DMEM and if I starve the cells with DMEM + 0.05% FBS, they look very bad the following day and lots of cells have died. But if I starve with DMEM + 1 % FBS + neuregulin + forskolin, then the survive.
Any thoughts? Is this unusual?
Which kind of DMEN are you using?
I was trial different types of DMEN, and the best is Gibco™ DMEM, high glucose, GlutaMAX (#11574516).
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