Hi,

I have been trying to establish primary BMDM from C57 mice. We have consistently found our macrophages are CD11b+ F480+ CD11c+. In our protocol, we seed cells at 0.5mil cells/cm2 (4.75 mil cells per well of 6-well plate) and MCSF (10ng/mL) is added on Day1. We have full media change on Day 3 and replenish the cells with MCSF and media. We harvest cells with accutase on day 7. The morphology does not look like M1 though. I saw a paper mentioned about high seeding density may affect CD11c expression. I felt our seeding density is quite high. Can anyone provide some explanation or suggestions? Thanks!!!