Posted 21 October 2009 - 10:46 PM
I am working with a cell line which is terribly clumpy. It is maintained on fibronectin-coated dishes and there are two types of populations: floating clumps and adherent ones. Both types are equally sticky/clumpy after digestion with trypsin-EDTA (Invitrogen's). In fact, I could see the single cells moving towards each other and forming doublet/triplets and more, within 10 mins of post-flushing with medium+FCS.
Any suggestions as to how I can successfully maintain a single cell suspension long enough for sorting?
Posted 25 October 2009 - 09:51 PM
Can you tell us the name of the cell line?
errr, my boss prefers to keep it anonymous for now as most of the data of this cell line is still unpublished .
Posted 25 November 2009 - 12:30 PM
Try to use up to 5 mM EDTA in your sorting buffer, usually it helps.