insect cell line problems

Carmen P Pena Diaz via methods%40net.bio.net (by C.P.Pena-Diaz from 2006.hull.ac.uk)
Tue Feb 23 06:46:42 EST 2010


Thank you all for your suggestions..... Have already started trying out some.

cheers

P.

-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu on behalf of Nikola Wenta
Sent: Mon 22/02/2010 17:41
To: methods from oat.bio.indiana.edu
Subject: RE: insect cell line problems
 
Hi Priscila,
first of all, in our lab we use Insect Xpress Medium (Lonza), supplemented with Pen/Strep, fungizide Partricine (you might try fungizone) and Pluconic (Gibco). We are doing them in round (not Erlenmeyer) shaking flasks (40 rpm) at 27 deg Celsius.
Sf9 cells usually stop growing when the split-rate is too high (after weeks, they recover anyway). You should count the cells in a hematozyte counter and then split them at density of ~8-12 million/ml (in our lab, it takes about 3-4 days) to not less than 1 million/ml. When you infect the cells, 2 million/ml is a good cell density. We do the shaking for virus amplification, protein expression and cell maintanance. Try it, and if you still have questions, :-)
Niko

Dr. Nikola Wenta
Research Fellow
D82
School of Biomedical Sciences
University of Nottingham
Queen's Medical Centre
Nottingham NG7 2UH
Tel: +44-(0)115-8230298 
This message has been checked for viruses but the contents of an attachment
may still contain software viruses which could damage your computer system:
you are advised to perform your own checks. Email communications with the
University of Nottingham may be monitored as permitted by UK legislation._______________________________________________
Methods mailing list
Methods from net.bio.net
http://www.bio.net/biomail/listinfo/methods

-------------- next part --------------
*****************************************************************************************
To view the terms under which this email is distributed, please go to http://www.hull.ac.uk/legal/email_disclaimer.html
*****************************************************************************************


More information about the Methods mailing list