separation of dead/alive T cell hybridoma cells

Paula Lavery plaver at PO-Box.McGill.CA
Wed Sep 25 09:54:22 EST 1996


>   bboverduin at ucdavis.edu (Bert Overduin) writes:
>  
>  I am looking for a method to separate alive and dead mouse suspension T
>  cell hybridoma (3DO) cells other than fluorescence activated cell sorting.
>  I am not very familiar with animal cell biological techniques, but guess
>  there must be a way to accomplish this. Answers and or hints can dbe
>  directly sent to me or posted on the newsgroup.
>  
>  Bert Overduin
>  CEPRAP / UC Davis
>  bboverduin at ucdavis.edu
>  
>  
>>>>
Try spinning your cell suspension over a Ficoll-Hypaque gradient.  I have successfully 
done this to clean up both hybridoma cells as well as preparations of activated cells.  
Cells harvested from the interface are usually 95-100% viable.
You will probably lose some cells in the process, so if cell numbers are crucial you may
want to try another method first.

Good luck!

Paula Lavery
Transplant Immunology Lab.
Royal Victoria Hospital/McGill University



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