How to seed adherent cells evenly in 150mm dishes? - (Jul/23/2007 )

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QUOTE (genehunter-1 @ Aug 6 2007, 10:55 PM)

I found the easiest way to do this is to add suspended cells to 30-35 ml medium in a 50 ml centrifuge tube, mix well, pour it into the plate and place the plate in a LEVELED incubator. It has been working very well for me.

How much medium do you use in a 150mm plate?
I use 20ml.
When transferring medium, I always use serological pipette, I don't "pour".
I'm afraid pour will cause cantamination. I never dare to try it.
Actually I handle at least 10 150mm plates at the same time:
I use them to culture AAV293 cells and produce AAV.
What do you use 150mm plates for?

-Bradley-

25-35 ml/plate.

I like the simplicity of this method, particularly when I work with multiple plates.
I have been using it for many years with little problem. Believe or not, accidentally touching the tip of a pippet and use it unknowingly afterwards could be the single the most frequent source of contamination. This method in principle can reduce the chance of getting contaminated. I usally spead some 75% EtOH on the outside of the bottle after use. I really have had no problem with this method.

I did adv production with 293 cells before. The medium turn yellow after 48 hrs. So I prefer adding more medium than less.

----A kind note: please be kind to people like scolix and anyone else who are willing to spend their time trying to help each other. We are fortunate to have them here. After all, they dont get paid for doing this.

Right? guys?

cheers. :-))----

-genehunter-1-

QUOTE (genehunter-1 @ Aug 8 2007, 08:45 AM)

----A kind note: please be kind to people like scolix and anyone else who are willing to spend their time trying to help each other. We are fortunate to have them here. After all, they dont get paid for doing this.

Right? guys?

cheers. :-))----

Sure

Cheers

-Bradley-

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