re sorting CHO cells

From: D. Robert Sutherland (rob.sutherland@utoronto.ca)
Date: Thu Dec 12 2002 - 21:33:09 EST


Hi Paul,

although we have not sorted CHO cells post detachment nor tested
VE-cadherin, we have recently had success detaching the CHO cells from
culture dishes using a fairly new product called Accutase from Pheonix
Flow Systems.  The active enzyme(s) in this product are not disclosed
(see their web-site for pdf of this product) but in a preliminary
experiment we obtained a single cell suspension that exhibited very high
viability at all concentrations tested.  In one parallel experiment
using the primitive hematopoietic cell line KG1a, we noted that surface
expression of CD34 (class III epitope), CD109 and CD45 (J33, pan CD45)
were not cleaved at concentrations sufficient to effect detachment of
CHO cells. Might be worth a try.

Rob Sutherland
University Health Network
Toronto



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