viability and MACS separation purities

Ann Atzberger Ann.Atzberger at EMBL-Heidelberg.de
Fri Oct 22 08:02:18 EST 1999


>Hi,
>
>  One of our clients asked me the effect of lower viability on the Vario
>MACS separation purity.  Since antibodies nonspecifically bind to dead
>cells, is this something to worry about in say a population of cells that
>has an 80% live count after thawing? Is it necessary to run a ficoll
>gradient to enrich for the live cells before MACS purifying? What is an
>acceptable live count before proceeding?  I would like to hear from anyone
>with experience along these lines.  Thanks,
>
>Kristi R. Harkins, PhD
>Director, Cell & Hybridoma Facility
>Iowa State University
>1104 Molecular Biology Building
>Ames, IA 50011
>Office (515) 294-2472
>Lab    (515) 294-8504
>FAX    (515) 294-0453
>http://www.biotech.iastate.edu/facilities/cellhyb/homepage.htm


Hi Kristi,

didn't quite understand the query.

Are you saying you're getting poor viability wth the MACS??? What kind of cells?

With 80% viability before sorting which is good- I don't see a need for Ficoll.

I haven't ever seen much non-specific binding to dead cells; but this could
be an issue if the population you are trying to isolate is very small.

My own experience with the MACS system for various types of cells is good
for recovery, viability and enrichment.

Check out the MACS web page, they also have a discussion forum.

Ann






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