monocyte devired macrophage - (Aug/21/2009 )
Hi,
I try to get monocytes derived macrophage. Monocytes were isolated from PBMC by ficoll and CD14 Microbeads. Monocytes were cultured in DMEM with M-CSF, heat inactivated human serum,ciprofloxacin and geatamicin. After 2 days stimulation, I found there are still a lot of unattached cell(I seed cells to 24 well plate at 1.2*106/well ). I thought they should turn to adherent cells after overnight stimulation and be fully differentiated at 7 days culture. I wonder how long monocytes should completely attach to plate?
Thank you very much for any suggestion and reply
-virus-
virus on Aug 21 2009, 09:20 AM said:
Hi,
I try to get monocytes derived macrophage. Monocytes were isolated from PBMC by ficoll and CD14 Microbeads. Monocytes were cultured in DMEM with M-CSF, heat inactivated human serum,ciprofloxacin and geatamicin. After 2 days stimulation, I found there are still a lot of unattached cell(I seed cells to 24 well plate at 1.2*106/well ). I thought they should turn to adherent cells after overnight stimulation and be fully differentiated at 7 days culture. I wonder how long monocytes should completely attach to plate?
Thank you very much for any suggestion and reply
I try to get monocytes derived macrophage. Monocytes were isolated from PBMC by ficoll and CD14 Microbeads. Monocytes were cultured in DMEM with M-CSF, heat inactivated human serum,ciprofloxacin and geatamicin. After 2 days stimulation, I found there are still a lot of unattached cell(I seed cells to 24 well plate at 1.2*106/well ). I thought they should turn to adherent cells after overnight stimulation and be fully differentiated at 7 days culture. I wonder how long monocytes should completely attach to plate?
Thank you very much for any suggestion and reply
We have tried TPA-induced differentiation of U937 premonocytic cells. It works with the doses like 10-30 nM and 50% cells approximately differentiates withing 2 days..
-Thapa-