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
errr, my boss prefers to keep it anonymous for now as most of the data of this cell line is still unpublished .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.