separation of two types of cells, eipithelial origin and ;ymphocyte origin

Robert Hartley rh at mblab.gla.ac.uk
Tue Dec 5 21:29:58 EST 2000


In article
<Pine.GSO.4.21.0012050957390.17470-100000 at inch.interchange.ubc.ca>,
lesley at interchange.ubc.ca (Lesley Weston) wrote:


Yep this is a good way of doing it before then pulling down with something
like dynal beads.

Remember always to then do  an Ab check for markers on sparse clones

Cheers
Bob

One thing not to do is grow the culture to confluence as epithellial and
endothellial will excise thmselves as rounded cells in the middle of a
clone and come out with the lymph's in Lesley's method

Cheers
Bob; Rainy Scotland


> Differential trypsinisation might work - it does on epithelium and
> fibroblasts. Use trypsin in citrate saline, with no EDTA, and leave it on
> a very short time, until some of the cells have detached. You can check
> progress every minute or so. With luck, one type of cells will detach
> before the other. When you see some loose cells, quickly quench the
> trypsin with medium + serum, remove the liquid and plate it into another
> dish or flask. Add medium to the original. You might have to do this more
> than once, but after 2 or 3 rounds, you should have 2 pure cultures. Hope
> this helps.
> 
> Lesley Weston.
> 
> 
> 
> On Tue, 5 Dec 2000, taro kijima wrote:
> 
> > Does anybody know how to separate two types of cell cultured in the same
> > flask. One is adhesive cells(epithelial origin). Another is lymphocyte
> > origin. We have to do PCR using cells after separating two types of cells.
> > 
> > 
> > 
> 
> 
> ---

-- 
Robert Hartley, Centre for Cell Engineering,Joseph Black Building
University of Glasgow, Glasgow G12 8QQ Tel: +44 (0)141 330 4756, 
Fax: +44 (0)141 330 3730 mailto:rh at mblab.gla.ac.uk
Web : http://www.gla.ac.uk/Inter/CellEngineering






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