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transfection of suspension typecell


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#1 anonymous

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Posted 12 February 2001 - 10:00 PM

Hi! I am trying to transfect N1S1, the suspension type cell line by cathionic lipid complex. till now, I have worked with adherent type cell lines. so I have no idea how I do same thing with suspension type cell. If any of you have ever transfected the suspension type cell with any methods, please give me some comments. Thank you for any suggestion!



#2 anonymous

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Posted 13 February 2001 - 10:00 PM

Hi, I've been doing some transient transfection experiments on human monocytic U937 cells which are basically suspension cells. Among all the approaches I've tried including liposome-mediated, DEAE-dextran, and electroporation, the use of FuGene6 agent from ROCHE has shown the best efficiency in transfection plus the greatest cell viability (>80%) because you could add serum to the medium during transfection. I don't know if it also works well with your cell line, but you could definitely try it. Also, to my experience the suspension cells prepared for transfection should not be passed beyound 4-5 generations otherwise the transfection efficiency will be somewhat reduced due to the overgrowth of the cells. Good luck to your research.

Jason


#3 anonymous

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Posted 09 March 2001 - 10:00 PM

Hi,Kim, there are two alternatives.One is centrifuge the cells before transfection, then resuspend the cells into the transfection reagent( in your case, mixure of DNA and liposome ), directly leave the centrifuge tube in RT or required temperature for sometime.

The other method you might try is to use collagen or some other materials which can allow the suspension cells attach to the flask.

Wish you success.


#4 anonymous

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Posted 19 July 2001 - 09:00 PM

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