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Help with HELA and HEK293 cell seeding for transfection


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#1 losybelle

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Posted 29 April 2011 - 07:50 AM

Hello everybody:

I need some help with my HEK and HELA cultures. I need to do some transfections in 24 and 6 wells plates, the problem is I cannot find the right amount of cells to seed of any culture in any tipe of plate. I tried around 125.000 cells for HELA 24well (my labmate told me to) to try to have around 50% confluency next day morning, but i got almos 80% (i need 30%-50% to sirna transfections).
But with HEK293, i tried 300.000 cells in 6well plate, and they were too little 24 hours later.

I know that i should keep trying till i find the apropriate ones, but i don't have so much time (i need to do all my experiments n a couple of months). Do you know how many cells should i use or a least where can I find any table or list Size/cell quantity?

Thank you very much

#2 chimpsarehungry

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Posted 29 April 2011 - 07:54 AM

I use 1.5 million HEK293Ts in a 6 well plate for 50-70% confluency by 24 hr later

#3 chimpsarehungry

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Posted 29 April 2011 - 07:56 AM

Or 1/6th of a confluent 100mm plate of 293Ts into a 60mm plate for 70% confluency about 18 hours later

#4 losybelle

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Posted 29 April 2011 - 12:20 PM

I use 1.5 million HEK293Ts in a 6 well plate for 50-70% confluency by 24 hr later


The 1.5 million is for the 6 wells right?

Thank you very much!

#5 chimpsarehungry

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Posted 29 April 2011 - 12:55 PM


I use 1.5 million HEK293Ts in a 6 well plate for 50-70% confluency by 24 hr later


The 1.5 million is for the 6 wells right?

Thank you very much!





Yep. Youre welcome






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