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About R28 cells culture


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3 replies to this topic

#1 iamfat

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Posted 10 September 2004 - 11:41 PM

It seems that R28 cells(a kind of retinal cells) without transfection have G418 resistence?
Have anyone met the same problem?

#2 kant0008

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Posted 12 September 2004 - 04:56 PM

Hi,
how much G418 are you using? They could have some "natural" resistance, depends on what's your dose?
Also, if they have been sitting in culture side by side with transfected lines- unlikely but possible (happened in out lab) cross-contamination can occur. Check for plasmid by PCR?
Also, someone earlier had the same problem with cells that they got from someone else- maybe you don't have all the info?
Hope this helps

#3 iamfat

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Posted 13 September 2004 - 06:58 AM

I cultured them for G418 death curve firstly without any plasmid transfected. G418 cannot kill those cells even in a concentration of 1400ug/ml!
So the only reason perhaps is that the cell is a gift from other labs. :)

#4 gseigel

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Posted 06 March 2012 - 09:57 AM

Hi everyone, I stumbled onto this question in a Google search. I am the PI who created the R28 cells. If you go back to our original papers, you'll see that these cells are already G418 resistant due to the immortalization process with the 12S E1A gene. I am very careful about sending these cells directly from my lab, so if you've gotten them from someone else, it was not the way that I would prefer. I always send an instruction sheet with the cells and their characteristics. I realize that this original question is from 2004, so hopefully you were able to figure this out. But I wanted to respond so that others could potentially benefit.



Establishment of an E1A-immortalized retinal cell culture.

Seigel GM.
In Vitro Cell Dev Biol Anim. 1996 Feb;32(2):66-8.




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