In the lab we used to use Matrigel (10 µl) for coating of 8-well Ibidi chambers and added cells with DMEM (Gibco 11965092) + 5% Matrigel. This worked fine but if we wanted to do imaging you needed to be very carefull during washing steps and especially after fixation with 2% PFA because the Matrigel starts to dissolve. Since we wanted a more user friendly approach we switched to do laminin coating (1:10 in DMEM, for 1h at 37°C). This worked fine until we needed to change the DMEM from Gibco to Capricorn (DMEM-HHSTA). Its only difference to Gibco is that it contains 25 mM HEPES.
Now all cells cysts are inverted and we have a lot of 2D cell batches.
Is really the media the problem? In 2D they grow fine.
Or might it be that it is just a bad laminin batch?
Also does anyone now a better protocol to fix and stain cysts grown on Matrigel coating?
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