From owner-dros@hgmp.mrc.ac.uk Tue May 14 21:53:39 2002 Return-Path: Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 983C617A80 for ; Tue, 14 May 2002 21:53:38 +0100 (BST) Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 144BD17C17 for ; Tue, 14 May 2002 21:53:34 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6020) id 8BAA917A80; Tue, 14 May 2002 21:53:32 +0100 (BST) Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 7651417C17 for ; Tue, 14 May 2002 18:06:08 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 60001) id 3DE2A17BB6; Tue, 14 May 2002 18:06:06 +0100 (BST) To: bionet-drosophila@net.bio.net From: rowan_greene@yahoo.com (Rowan Greene) Newsgroups: bionet.drosophila Subject: Re: S2 cell question Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Message-ID: <20020514170556.20713.qmail@web13108.mail.yahoo.com> MIME-Version: 1.0 Content-Type: text/plain; charset=us-ascii X-Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id DCEFE17C1C for ; Tue, 14 May 2002 18:06:01 +0100 (BST) X-Received: from web13108.mail.yahoo.com (web13108.mail.yahoo.com [216.136.174.153]) by mercury.hgmp.mrc.ac.uk (Postfix) with SMTP id 8ED0D17BB6 for ; Tue, 14 May 2002 18:05:57 +0100 (BST) X-Received: from [130.132.227.110] by web13108.mail.yahoo.com via HTTP; Tue, 14 May 2002 10:05:56 PDT X-To: dros@net.bio.net Date: Tue, 14 May 2002 21:53:32 +0100 (BST) Sender: owner-dros@hgmp.mrc.ac.uk Precedence: bulk I have just started working with S2 cells (actually TC in general). I have been trying to thaw cells but with no success. These aliquots have been used successfully fairly recently. It looks as though the cells are not attaching to the flask and are not reaching confluency. The protocol for this that I have been following is quick thaw of a 1.5 mL aliquot, transfer to 30 mL complete media, passage after 3 days because of clumping. The cells I have thawed in this manner are still not confluent after 1.5 weeks. I have tried this several times now. This simple method of thawing and not washing out the DMSO has been used routinely in the past in our lab successfully. Do I just need to be patient with these cells (ie giving them more time to acclimate)? Is there a reason they aren't attaching? Any suggestions are appreciated. Thanks. Rowan Greene __________________________________________________ Do You Yahoo!? LAUNCH - Your Yahoo! Music Experience http://launch.yahoo.com --- .