I am working with human neurons in vitro and want to start an experiment where I can co-culture neurons with M1 macrophages. RPMI is deadly for neurons. Serum is not compatible for pure neuronal cultures. I grow my neurons in DMEM/F12 supplemented with growth factors to support neuron survival. To start the experiment of co-culture, I would like to know if DMEM/F12 medium without serum can be compatible for macrophages (after polarization and activation as M1)? and how long the macrophages can survive in such conditions?
Any comments?
Thanks a lot in advance,
Laila :)
They can survive more than couple weeks, but not sure about functions.
From what source you have isolate macrophages?
Thank you for your reply. I will isolate/polarize them from human blood monocytes
You believe they can survive for 2 weeks without serum? That would be great. I will use a uni-culture controls (macrophages or neurons alone) to see the difference in function. Do you have any recommendation at what functions I should look for to tell me that macrophages have changes or not?
Thanks again :)
I'm working with peritoneal macrophages and bone marrow stromal cells. Both of mentioned primary cultures are mix of the different cells including macrophages.
Without serum attached cells can survive at least 2 week, but foam cell formation could be observed (serum was at first step, but monocytes contacted to serum also).
Please follow these discussions:
https://www.researchgate.net/post/Serum_free_medium_for_monocyte-macrophage_differentiation
https://www.researchgate.net/post/Human_Monocyte_Derived_Macrophages
And these articles could help:
https://www.researchgate.net/publication/21637312_Monocyte_proliferation_in_a_cytokine-free_serum-free_system
http://www.jleukbio.org/content/44/2/111.full.pdf
http://core.ac.uk/download/pdf/11544508.pdf
Article Monocyte proliferation in a cytokine-free, serum-free system
Thanks a lot guys for providing suggestions, info and references.
Laila :)
As alternate you can try special serum-free culture media from Lonza. Works perfect with primary monocytes/macrophages from peripheral blood
Thanks Olga. I am trying X-VIVO15 serum free medium with both neurons and monocytes/macrophages. I can let you know if it works, if anyone is interested.
Good luck all :)
They can live for 2 weeks, but you need to consider cell activity, so I suggest that no more than 2 days
hi Laila, can you let us know if X-VIVO15 serum free medium with monocytes/macrophages worked well? how did you check that it worked on BMDM? thank you!
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