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Insect cell sticking to glassware in suspension culture

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Insect cell sticking to glassware in suspension culture

Postby imm » Thu Mar 24, 2005 4:55 am

Hi
I have been growing and expressing proteins in insect cells for aprox 4 years. When I grow my cells in shaker cultures, I get a 'sludge' ring of cells that stick to the glass flask, just slightly higher than the media level.
After a few hours of placing cells in a new flask, I can see the start of this cell loss. As these cells are not in contact with the media they die.
From this I have 2 problems
1. Cell viabilty drops
2. A lot of the cells I seed into the flask end up on the glassware, so my cell number is much lower than it should be. (as a result I am getting very low expression of my protein since not enough cells in culture).
This problem seems to have gotten worse and I have afeeling it is related to the way I am growing the cells (no mycoplasma and have this problem with Sf21 and High 5 cells).

Anyone know what might be happening ??
Thanks in advance
imm
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Postby biostudent84 » Thu Mar 24, 2005 5:09 am

Did you try manually stirring the solution every 15 minutes or so?
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Postby imm » Thu Mar 24, 2005 6:08 am

Thanks for reply but I leave these cells culturing 24-7 so would be impossible to manually stir at this interval
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Postby biostudent84 » Thu Mar 24, 2005 6:35 am

That's the only thing I can think of...perhaps if you and two other people take "shifts" to stir? I'm afraid that the motions of any automated shaker/stirrer would also give you this problem.

LOL, sometimes, in the interests of carrying out a scientific expirement requires a little sleep deprivation...
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Postby DevGrp » Thu Mar 24, 2005 4:32 pm

How about siliconizing the glass with something like Sigmacote (Sigma)?

It might help.

Ian
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thanks

Postby imm » Sun Mar 27, 2005 6:56 am

Thank you both for your help, will try the Sigmacote :)
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