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New patch of sf9 cells not growing


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#1 saad

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Posted 04 June 2012 - 12:34 AM

Hi all,

I've gotten a new batch of adapted to suspension sf9 insect cells from Invitogen. I use SFM-900 II at 27 degree, 120 rpm, no serum or antibiotic at all.
It is now 4 days and the cells seem not grwoing. I counted the cells after 2 days and they were 0.7 million/mL but after 4 days the same or less with no noticeable dead cells.
Is it possible for the new cells to take longer time to grow I mean for the first time.
I follow the manufature's instructions. Thawed the vial quickly and add to pre-warmed 27 mL media.
Any suggestion or recommendation will be highly appreciated.
The 1 mL cells cost more $450.!!!

Thank you so much for you help.

#2 protolder

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Posted 16 June 2012 - 01:50 PM

Hola,I had similar problems with Mimic Sf9 cells , but the cells arrived dead.In your case, in which cells are living, I would seed in a B25 flask and let form monolayer in sf900. After, pass to a B75 scraping monolayer or pumping medium with a pipette, and when the density was enough to seed in suspension I will do it. This cells are able to give 5x10e6 cells/ml. Sorry by the delayed answer. Buena suerte

#3 saad

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Posted 17 June 2012 - 04:09 AM

Thank you Protolder,
Actually, I was very patient with this cells for 10-11 days untill they reached 1.9 x10^6 cells/mL.
After I passage them the first time they seem growing better. Now I'm at passage 4 they are doubleing each 24-48 hours.

Thnank you again,




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