insect cell line problems
(by Nikola.Wenta from nottingham.ac.uk)
Mon Feb 22 12:41:33 EST 2010
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, :-)
Dr. Nikola Wenta
School of Biomedical Sciences
University of Nottingham
Queen's Medical Centre
Nottingham NG7 2UH
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