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monocyte devired macrophage


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#1 virus

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Posted 21 August 2009 - 08:20 AM

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

#2 Thapa

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Posted 21 August 2009 - 10:14 PM

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



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..
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