experience sharing: the whole cell of 293 T . I hope to be helpful

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zhangxu04
zhangxu04's picture
experience sharing: the whole cell of 293 T . I hope to be helpful

    I have done patch clamp with 293 for 2 month. it was said the 293 was very eary to patch. but I meet much trouble and think out many method to  solve these trouble. but now I found I made a mistake in choosing the cell.
I compared 239A with 293T. I found 293A was to thin and fragile to patct. and alway I sucked the 293A into the tip. while 293T was more easy  to form whole cell. I read some paper the 293T was the cell line that should be choosn to be transfected. in fact I waste some of my time. I hope my experiemce will be helpful.
also I hope you can give your suggestion and opinion about the difference of this two cell line espically the electrophysiological related difference.
 

Fraser Moss
Fraser Moss's picture
293T cells constitutively

293T cells constitutively express the T-antigen which allows episomal replication of plasmids that contain the SV40 origin of replication such as the commonly used pcDNA3.1.
This means that the daughter cells which arise from cell division after transient transfection are able to replicate the plasmid and have high expression levels of the tranfected cDNA like the primarily transfected cells.
293/293A do not express the T-antigen, so as your cells divide, the expression levels in each cell become diluted.
Beware though there are often circumstances when you do not want to massively over express your gene of interest, and in these cases 293T cells may not be suitable for your study.
also For Your electrophysiological information, in my hands HEK293Ts (from ATCC CRL-11268 have an endogenous outward rectfying potassium current).