Hi,
I'm trying to establish a co-culture of decidua cells and peritoneal Macrophages, and my question is how can I sort these two different cells after doing the co-culture? (Since these two kinds of cells are adherent to petri dish substrate) and I was wondering is there a way besides MACS to separate the cells because I'm trying to use a procedure that is more accurate and economical.
thanks in advance.
Dear Nasim!
Maybe try this protocol
Article Harvest and Culture of Mouse Peritoneal Macrophages
12. Centrifuge peritoneal cells (300 x g; 3 min) and collect cell pellet. Usually about 2-4 million resident peritoneal cells can be recovered from one C57BL/6 mouse using this method and about 50% are peritoneal macrophages.
13. Resuspend cell pellet from one mouse in 1 ml of cRPMI medium, count cells. 14. Culture peritoneal cells in a 12-well plate, 2 million/well in 1 ml cRPMI at 37 °C with 5% CO2 for 6-18 h. During this time, peritoneal macrophages adhere to the plastic surface. The floating non-macrophages can then be washed away by adding and aspirating 0.5 ml cRPMI medium twice.
Alexandr Chernov
I really appreciate for your answer Mr. Chernov but as I mentioned my exact question is how can I separate two kinds of cells (decidua and peritoneal macrophages) after a co-culture besides MACS ?
Hi Nasim,
There is an old technique termed panning. Essentially how we did this was buy antibody conjugated dynabeads which we incubated with cells (in sunspension) at 4 degrees and then pressed the resulting mixture against a large magnet to pull out the macrophages. You will want to use an antibodyconjugated bead that is sovific for mouse macrophages
As mentioned by Lora, panning, and flow cytometry can be used.
https://doi.org/10.1002/eji.201646632
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