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transfection of suspended CHO cells

transfection reagent

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#1 sara.r

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Posted 22 July 2015 - 11:43 AM

Hi

I am trying to transfect CHO-S cells in static culture and I have used lipofectamine 2000 and lipofectamine LTX transfection reagents (I do not have access to an electroporation device). I use a GFP containing reporter plasmid and I analyze GFP expression 24-72 hrs post transfection using fluorescent microscopy. I have tried different reagent:DNA concentrations but I have not have any success in transfection. I was wondering if anyone has any experience in transfection of suspension adopted cells.

 

Bests



#2 bob1

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Posted 22 July 2015 - 12:51 PM

Suspension cells should be (in theory) more transfectable than attached cells as all the surface area of the cell is available for contact with the transfection reagent.

 

There are a few things you could try - the most obvious is to transfect another cell line such as HeLa or HEK293, that you know are easily transfectable, so that you can test that the GFP expression is working, and that you can do a transfection with these reagents.

 

I have heard that CHO are relatively hard to transfect, but haven't actually worked with them for transfection purposes myself.



#3 sara.r

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Posted 23 July 2015 - 03:03 AM

Hi

Thanks for your comment. I have actually transfected CHO-K1 (adherent cells) with the same reagent and the same reporter plasmid and the majority of cells were transfected (more than 70 percent).






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