I am working on CHO cells and am trying to come up with an alternate method for trypsinizing the adhered cells. Since my study is about estimating the activity of insulin receptors, I am afraid trypsin would cause a permanent activation of insulin receptors which is undesirable in my case. I am losing a lot of cells whilst scraping and I found it to be an inefficient alternative for study on whole cells (I wouldn't worry too much about scraping when homogenizing and solubilizing the membrane).
Does PBS-EDTA (without MgCl2 and CaCl2) help? Can anyone kindly suggest to me your ideas?
Thank you.
By the very definition, it is impossible to "trypsinize" anything without trypsin, an enzyme! ;)
That said, PBS-EDTA/EGTA washes can help with some cells that weakly adhere to plates (not sure about CHO).
Your best option will be to consider growing them in suspension as non-adherent cells, if you need intact and live cells for your studies. There are adaptation protocols available for such switches.
You could try scraping them into PBS if you plan to lyse them afterwards. Not sure for splitting cultures.
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