Sf9 Cells

Michael CHENG a52041 at sakura.kudpc.kyoto-u.ac.jp
Sat Aug 29 01:07:01 EST 1992


  Hi! I was wondering if anyone out there has experience working with Sf9 cells
isolated from the ovary of armyworms.
  I pretty much follow the protocol of Summers and Smith. Using Grace's medium
supplemented with 10% FCS and the specified concentration of Gentamicin.
  However, recently my cells begin to die after a few days of incubation.
Specifically, after reviving the cells from liquid N2 and seeding 2e6 cells in
a 25cm2 flask, the cells are fine and reaches confluency. I would then pass the
cells 1 to 3 or 1 to 4 in about 5 days. But appoximately 2 days after the 1st
passage, the cells would begin to die. They would float, stop dividing, and I
think that I can see holes in them (some bright spots?). Don't think it's
contamination by baculovirus though that's what I'm growing the cells for?
Has anyone experienced this? Any suggestions?
  Are these cells susceptible to mycoplasma contamination? The gentamicin that
I'm using is from BRL and the maker describes as effective against mycoplasma.
  I've also heard that a "new?" medium called Sf900 is better. Any comments?
   How many passages can I achieve with these cells continously
  Right now, I plan on purchasing new cells from INV.
  I would appreciate any comments or suggestions.
  Thanks in advance.

a52041 at sakura.kudpc.kyoto-u.ac.jp
Michael Cheng
?



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