Hi,Thank you for posting this protocol. I am starting to work...

protocol Protocol: Isolation and Culture of Mouse Bone Marrow-derived Macrophages (BMM’phi’)
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Thank you for posting this protocol. I am starting to work with BM-derived macrophages and have followed other protocols, and I have been getting impure cultures where I see fibroblasts-like cells as well as round cells that I believe are monocytes/macrophages. In these case
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yu liming Answered Aug 30, 2013

Nanjing Medical University

Hi !
I gotta a question about Procedure 3(g), how could you eliminate erythrocyte and fragments of bones? I usually use red cell lysis buffer as well as filter with 200 nylon mesh , but the rate of adhesion is very low . I am confused about that.
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Ran Chen Author Answered Sep 3, 2011

Zhejiang JFK Biological Technology Co. Ltd.

L929 conditioned medium is an alternative to M-CSF. The protocol above is to get as large number as you can. If you want highly pure macrophages, following the step below may help:

During the culture of BMM cells, wash cells twice with PBS every 2 days, and then change to fresh BMM growth medium. -- Macrophage progenitors adhere to the plastic firmly and are not washed away. Fully differentiated macrophages can be harvested and used at day 7.
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