Thioglycollate induced inflammation in mice

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A.N.
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Thioglycollate induced inflammation in mice

i want to get neutrophils out of the peritoneum after an injection of 1ml 5% thioglycollate solution. I removed the cells with 5ml 30% sucrose 6h after the injection. The problem is, the cell number 6h after the injection is not different to the cell number of uninjected mice. So the inflammation did not work?
Has anyone experience with this thioglycollate model and can give me a tip?

Tracy
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http://jimmunol.org/cgi

http://jimmunol.org/cgi/content/abstract/141/6/2090

You can check this paper for the details of this method.

I don't have experience in this.

Jen_Floyd
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I thought it was mainly

I thought it was mainly macrophages elicited by thioglycolate.

A Timmer
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There should be an early

There should be an early influx of neutrophils at the site of injection. We use 3 ml of 3% thioglycollate to recruit immune cells. It is also possible that your thioglycollate isn't that great. At least for macrophage collection, I have noticed huge variations in numbers depending on the batch of thio that I used. I have also heard that after making and autoclaving the thio, it is best to let it "age" for a while before using and that gives the highest yields.

Jen_Floyd
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Yes, it should be aged for

Yes, it should be aged for maximal effect. Our batch was brought over by someone when she moved from another lab.

Lim
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I have been using 5%

I have been using 5% thioglycolate to induce peritoneal macrophage in mouse. But the solution colour turns to cloudy instead of dark blue. I found this able to induce high amount of macrophage. I am not sure what is the principal behind this.

heehawmcduff
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We have used 1ml of 5%

We have used 1ml of 5% solution in the past for recruiting macs but you will definitely get an early neutrophil influx with it.  You could also try LPS as a positive control if required.

rajeevnayan22
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To Lim,  Your Thioglycollate

To Lim,  Your Thioglycollate solution turns cloudy because of contamination. U should not use it cause it makes no sense of inducing inflammation with bacterial contamination. Ya, definitely it will induce strong inflammation cause of the bacterial contamination.