Culture of NIH3T3-CD40L
Posted 09 August 2009 - 11:51 PM
I'm new to immunology and now culturing NIH3T3-CD40L for proliferating B cells.
However, I meet serious trouble in culturing NIH3T3-CD40L cells as I found that the NIH3T3-CD40L cells detached from 6-well plate (TPP) seriously after seeding B cells (PBMC) for 2 to 3 days.
I also observe that there is a change in the growth rate of the NIH3T3-CD40 cells. At first, I seed 0.4 x 10^6 NIH3T3-CD40L cells to a well and it can confluent on the next day. However, it took two days to confluent after several passage of NIH3T3-CD40L cells. I seed 2 to 3 x 10^6 cells in one T75 flask and took two days to confluent. I wonder if there is anything wrong during the passage of 3T3 cells??
Is there any tricks or tips in culturing NIH3T3-CD40L cells? Will the use of Corning 6-well plate better than TPP 6-well plates?
Please kindly help!
THANK YOU VERY MUCH!!!
Posted 10 August 2009 - 05:22 PM
Posted 10 August 2009 - 10:36 PM
Thank you very much Bob! I'm trying to thaw a new cell line of 3T3 and lowering the density. Is there any optimal density for culturing 3T3? In addition, how can mycoplasm be checked? I heard from other lab that mycoplasma contamination is difficult to be detected.
Posted 11 August 2009 - 04:35 PM
Posted 12 August 2009 - 10:16 PM
Thank you very much!! I will try to check the contamination!!
Posted 27 April 2010 - 02:16 PM
Thanks for your help,