I grow organoids of ovarian cancer cell lines A2780 and SKOV-3, I embed them in matrigel...after reaching confluency I need to perform a Western blot, but I do not know how to harvest the cells
Thank you Doctor.....do you use them at 37 degrees C or is it ok to use at 4?
Hello Rawda Elshennawy
The protocol that is used to harvest organoids embedded in matrigel depends very much on the type of protein you are interested in. Dispase as mentioned by Debasish contains proteases that could have a damaging effect on protein of interest. I would suggest you incubate the matrigel containing the organoids on ice on a shaker for agitation for a few hours. Matrigel depolymerizes at low temperature and therefore help in cell recovery. For a complete harvest you could use recovery solution from Corning or BD. You could even try trypsin-EDTA treatment at 4°C if the matrigel does not depolymerize totally.
All the best.
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