I have been isolating PBMC's from feline blood with lymphoprep and it was all working fine. Now I still get good layers but when I centrifuge the isolated cells they clump and smear on the bottom of the tubes. We have just started using the new corning centristar tubes and have a new batch of FCS, both of these I will evaluate. Any other thoughts ?
are the clumps or smears breakable with pipetting???? did you check the viability? Cells are fine?
They are difficult to break up and sometimes just smear on the bottom of the tube. Viability and functio of the cells I get is okay. The yield has really dropped.
sorry for this late reply. but please check with lymphoprep too.
though it is clumping you can remove the smear and work with the remaining cells. If you carry further with some assay, keep a calculation in mind about the cells loss and take extra cells accordingly.
and also check blood collection quality and also breed of cats etc.
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