From: Annette Byrne (AByrne@pcyc.com)
Date: Tue Mar 04 2003 - 15:04:08 EST
Hi Flowers Does anyone have any advice on getting rid of debris from solid tumor suspensions ( derived by mechanical disaggregation) using a Ficoll gradient to sort debris from cellular matter. Also, I am having some difficulty in generating a reasonably live cell suspension for use on Flow, from A549 SQ tumors (about 2-300mg in size) grown in CD-1 nudes. I basically chop up the tumors, pass through a 100um mesh, spin down and filter again through a 50um syringe filter to run suspensions on Flow. What viability should I expect with this technique and does anyone know exactly how many cells one would hope to get out of a 200mg SQ non-necrotic tumor ? Thanks as always for any advice Annette Annette Byrne PhD Scientist Pharmacyclics 995 E Arques Ave Sunnyvale CA 94085-4521 USA Tel: 001-408-3283640 Fax: 001-408-3283689 e mail : AByrne@pcyc.com www.pharmacyclics.com
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