Monocyte culture - How to get the monocytes to adhere to culture plates? (Mar/06/2008 )

Hi,

I am trying to get monocytes to adhere to a tissue culture plate in order to perform an adhesion assays (by adding non adherent cells that express a receptor that should bind to the monocytes). Therefore, i need the monocytes to adhere quite strongly to the plate, otherwise they wash off when i remove the non adherent cells.
Right now, i cannot get them to adhere.... I am incubating PBMCs from human blood in RPMI with 10% serum for 2 hours, and was hoping this would be enough for them to adhere. I dont mind other cell contamination, i just want the monocytes to adhere, and as quickly as possible.
Any suggestions?

Thanks a lot

s.

coated the plates with poly-L-lysine or poly-L-ornthine.

It will do the trick.

by the way, the cells should adhere quite well even withput coating. have you checked cell viability ?

L.

Thanks,
I ll give it a try. Do you recommend media with or without serum?

Thanks

1. Regarding the PLL - make sure after you coat the pate (10ug/ml), remove the poly L-lysine with suction and leave to dry, wash once with PBS before planting the cells.

2. Serum - usually cells adhere faster without serum, but you should try it with your cells..

best of luck.

L.