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.
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.