I recently bought a LNCaP clone FGC (ATCC® CRL-1740™) vial and thawed it following all the steps described in datasheet. I used the recomended culture medium (RPMI-1640) and FBS at 10% (all from ATCC). However, the cells didn't attach. All of them are suspended in the medium. The cells are alive but they didn't differentiate or attach since I thawed (six days ago).
I already tried to change the flask (wondering if something was wrong with the plastic, but the response of the cells in the new flask was exactly the same, they are alive, suspended and indifferentiated). I also tried increasing the concentration of FBS in the medium to 20%, but nothing changed.
I would like to know if someone has any tips that can help me.
Hi:
These cells are very easy to detached from the flask. when you seed/subculture the cells, the flask should not be moved/shaken at least for 48 hours, otherwise, the cells detached and reseed again.
Hello!
Most likely, your cells did not survive the defrost. This culture is adherent, which means that the cells must adhere to grow. YOU Pop the cells with trypan blue to watch death
Did you centrifuge the cells and replace the medium in order to remove the DMSO before seeding the cells? I had problems with LNCaP not adhering as well, changing the culture dishes helped me. Maybe you need another type of dish. I use Falcon/Corning.
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