Hi,
Im trying to isolate intratestinal macrophages from mouse lungs, and grow them in culture for a few days, to collect secretome. Does somebody have a good protocol for it?
I tried face sorting but the cells didn't grow well and died in the following days.
I also tried to separate f4/80 cells with macs columns but didn't get any separation.
Can someone have any suggestions?
Thank you,
Shani
Hi: You may find this article very helpful:
https://dx.doi.org/10.1007%2F978-1-4939-7837-3_6
Mohammad Aqdas Thank you!
However, I tried flow sorting and for me the big problem in keep them in culture for few days.
Well in that case, you should try supplementing with r-MCSF or L929 supernatant in the cultures. Macrophages are kinda dependent on this growth factor. For more information, please see:
https://dx.doi.org/10.3791%2F57287
Good luck!