Can peritoneal macrophages grow?

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sxyan
sxyan's picture
Can peritoneal macrophages grow?

In my experiment, 3 days after 4% thioglycollate elicitation, i can not harvest too many peritoneal macrophage from mouse, about 2*106 cells per mouse. So i want to know can peritoneal macrophage grow more? Which reagent can be used to stimulate macrophages grow, GM-CSF or ?  Hope for your precious suggestions and thank you very much.

sxyan
sxyan's picture
i really want to know

i really want to know peritoneal macrophage can grow or not.
Can anybody give me a reply? 
Very thank you!

marcus muench
marcus muench's picture
Yes, peritoneal macrophages

Yes, peritoneal macrophages can grow to some degree.

Stewart CC, Lin HS, Adles C. Proliferation and colony-forming ability of peritoneal exudate cells in liquid culture. J Exp Med 1975, May;141(5):1114-32.

Peritoneal exudate cells, obtained from mice injected with thioglycollate medium and cultured in medium containing L-cell-conditioned medium, will proliferate in an exponential fashion for 18 days with a doubling time of 68 h. After a 2 h pulse of tritiated thymidine, labeled adherent cells increased to a maximum of 22-34% during the 1st and 2nd wk of culture. Increasing the cell concentration from 2 times 10-3 to 2 times 10-5 cells/culture reduced exponential growth to 10 days and the doubling time was increased to 81.6 h. Under these culture conditions, peritoneal exudate cells were shown to form colonies on the surface of culture dishes when plated at low density. The cells within the colony were shown to be macrophages using yeast and antibody-coated sheep erythrocytes as a test for phagocytic function. The plating efficiolonies arose from a single precursor cell. The adherent cell population contains the colony-forming precursors. These precursors can be stimulated to form colonies for at least 2 wk by the addition of conditioned medium to cultures at various times after plating. While very few colony-forming cells could be demonstrated in the unstimulated peritoneal lavage, their numbers begin to increase in the exudate 4 h after injection of thioglycollate medium and reach a maximum by day 3 and then decrease. Isolated colonies may be useful in studying the function of macrophages.

sxyan
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thank you very much for your

thank you very much for your reply and reference.  nowadays, i can isolate more peritoneal macrophage, and that's enough for further experiment. again, very thank you!

Noga Or-Geva
Noga Or-Geva's picture
How do you isolate many peritoneal macrophages?

I'm looking to isolate as many of the peritoneal macrophages as I can. I'm getting really low numbers using lavage and I need enought to be able to sort and then run expression studies. I do not want to inject anything proir to my treatment into the peritoneum because I want it to be clean. Any protocol? Any known way of blocking migration of the macrophages after my treatment?