which medium to use in cocultures - (Jan/10/2010 )
hello, i am culturing neuron and macrophage cell lines together in a co-culture. should i use the recommended medium for neurons or macrophages? or is there a separate medium i should use?
prof. moriarty on Jan 10 2010, 01:06 PM said:
Which cell lines specifically are you working with, and what are the recommended media? If they both grow in the same base media (DMEM, RPMI, etc.), I'd say just combine the supplements from the 2 media recipes. Also, have you checked the literature for precedent of a similar co-culture?
Well I know for certain that I am using the j774.2 macrophage cell line (dmem + 2mM l-glutamine + 10% fbs). as for the neuroblastoma cell line, i could either use the nd7/23 cell line (same medium as above) or n1e-115 (dmem + 4.5 g/L glucose + 10% fbs), or possibly another cell line if a fellow researcher is kind enough to donate a vial.
the nd7/23 cell line is obviously convenient, but it is a rat/mouse recombinant and seems to be mainly used for ion channel work. i'm not really sure if it's the best cell line for what i'm doing - basically i'm trying to find out whether inhibitors of a cytokine are neuroprotective.
the nd7 cell line has been co cultured with keratinocytes and other epithelial cells, but i haven't seen any literature for co culture with macrophages.
also, would you happen to know of methods for counting one cell line alone in co culture?
Looking at the HPACC catalog, it looks like all 3 cell lines should work with the same medium (DMEM+2mM L-Glut+10%FBS), so you should be fine in that respect.
As far as counting the cell types, if there's a major difference in size or morphology you could probably do it manually. Otherwise I'd say your best bet is probably FACS with staining for specific antigens. NeuN comes to mind for the neuroblastomas, and it should be easy enough to find a macrophage-specific marker.