HeLa S3 spinner culture

Alexandre Blais Alexandre.Blais at crchul.ulaval.ca
Wed Oct 27 07:19:17 EST 1999


Hi,

I have to purify an antigen found in human cells. Since this protein is
present in low levels, I have to prepare large amounts of cell extracts
to purify it. I decided to use HeLa S3 spinner cultures to reach my
goal. In our lab, we have some small and large spinner flasks, we have
the cell line HeLa S3 frozen in N2. But nobody has the knowledge of how
to grow this cell line in spinner flasks.I consulted Ian Fresney's book
but there are a lot of things that remain unclear.

I would like to know:

What is the medium I shoul use
What should I supplement it with (serum ? salts?)
Should I use anti-foaming agents
At which velocity should I set the stirring paddle
How do I set up the culture  (by beginning as an attached monolayer
culture, or by beginning directly as a suspension culture)
What are the minimal and maximal cell density allowed
And any other detail that is important

Thanks a lot for any hint

Alexandre Blais
Centre de Recherche du CHUL
Québec, Canada




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