help! 293F cell line

Nancy Hurtado-Ziola nhurtado at UCSD.Edu
Thu Jan 17 20:41:53 EST 2008

Hello fellow flow-ers,

I have a soluble protein that's really difficult to express.  I  
mentioned it to someone and they said I should use 293F cells to  
solve this problem.  Well, I need some information on these cells.   
I'm having trouble growing 293F cells.	I purchased them directly  
from Invitrogen.  The first batch started out OK.  I did everything  
the supplier said to do (use their SF media, use warmed media, keep  
cells at 37deg c, with 8% CO2 level, check viability with tripan  
blue, etc., etc.)  but the cell population diminished and there was  
so much debris.  I got to pass 3, but never had enough cells after  
that to do another pass let alone freeze down or for a transfection.   
I've never seen a photo of what these cells are supposed to look like  
after the culture takes off.  Do cells in suspension typically have a  
lot of debris?

I called Invitrogen and asked for help 3 different times during my  
attempt to grow these cells.  They finally sent me a new vial of  
cells.	Again, the cells started out ok.  The first pass, went ok,  
but now the cells seem to be dying and there is an increase in	
cellular debris. (I also thought it might be the media so I gave them  
the lot number, but there didn't seem to be any reported problems.)   
Has ANYONE seen this before?  If so, did you figure out what the  
problem was?  Were you able to resolve it and get a good culture going?

As always, I appreciate any help you might have to offer.
Nancy Hurtado-Ziola, Ph.D.
Varki Laboratory

Department of Cellular and Molecular Medicine
University of California, San Diego
9500 Gilman Dr.
CMM-E 1087, Mail Code 0687
La Jolla, CA 92093
Lab Phone: (858) 534-1346
Fax: (858) 534-5611

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