Do we have to use corn oil or olive oil?
It is necessary for corn oil has to be sterile?
Is there any protocol where pure CCl4 is used?
I need articles with IF>5.
I have used both and it worked fine.
You don't need sterilize the oil.
Adjust the optimal concentration for your aim.
I have read articles which oil is not used. We also used it once and, for our objective, it was ok.
Good Luck!
If you have any detailed protocol?
Can you read the full text of an article: http://www.ncbi.nlm.nih.gov/pubmed/25351459?
I unfortunately can read only the abstract.
Dear Jasmim,
i have this one, by my group. If you need a protocol, i think that this one its ok.
We have tried it with corn oil and it worked fine. We also tried a different protocol with a loading dose followed by maintenance doses for 5 weeks and it seemed that the effects of CCL4 was more tolerated by female more than male. The dose needs to be adjusted according to the body weight and longer protocol are safer.
Here are a few things that I’ve learned from my experience using CCl4 injections to induce liver inflammation and fibrosis in mice.
For the experiments that I was doing, the goal was to induce chronic inflammation/fibrosis – the dosage and injection regimen will be different (namely, a higher dose with fewer injections) if the goal is to induce acute liver damage.
Starting at 8 weeks of age, I injected male C57BL/6J mice intraperitoneally with a solution of 10% CCl4 in mineral oil. Mice were injected with ~2.5ul per gram body weight twice a week for 12 weeks. This protocol was sufficient to induce chronic liver inflammation, steatosis, and fibrosis that was maintained until at least 65 weeks of age.
I’ve tried both corn oil and mineral oil. In either case, with repeated IP injection the oil ends up forming small white inclusion bodies attached to the exterior of the peritoneal organs (liver, intestines, etc.). This is likely because the amount of oil injected is too much for the animals to completely process, so the excess gets sequestered in these inclusion bodies. (They are also formed with repeated injection of oil alone, so this is not an effect of the CCl4). In my experience, corn oil induced a stronger reaction in the mice, leading to a larger number and size of these inclusion bodies, as compared to mineral oil. As they are likely to affect the animals’ physiology in some way, my conclusion was that mineral oil was a more appropriate vehicle for delivery.
I would suggest using sterile oil – it may not be absolutely necessary, but its minimal expense is justified by the enhanced experimental control it provides. Sterile mineral oil is available from Sigma (product #M5310).
Given CCl4’s high volatility, I think it would be harder to maintain a consistent effective dose without mixing it into some sort of delivery vehicle.
I’d be happy to send a more detailed protocol and/or example images of what I described if you’re interested.
Good luck!
Jesse
Dear Jesse,
Of course I'm interested. Thank you very much.
Our team will start with experiment tomorrow by the following protocol (http://onlinelibrary.wiley.com/doi/10.1002/hep.27392/full):
CCl4 : corn oil = 1 : 3
1µl/g intraperioneally twice per week for 1 month.
I hope it will be good.
Greetings from the Centre for Molecular Medicine and Stem Cell Research, Serbia!
Jasmin
That protocol looks like a good one - it should work just fine. Here are a couple of images showing the inclusion bodies and Masson's trichrome staining of liver to show fibrosis (collagen stained blue)/steatosis. I've also included a more detailed protocol, although not everything will apply to your situation.
All the best!
Hi Jasmin,
in my research group we are doing a lot of experiments with CCL4, take a look at our published results. If any thing else you need just ask me...we did the acute models and chronic (not published jet)
Answers to other questions: olive oil, not sterile (but at 4 degrees kept), most def. pure CCL4 is dissolved in O.O. 1-2 hours before use.
all best
Nikola
Dear colleagues,
We are doing also some fibrosis model with CCl4 and corn oil 10% solution. We will inject it 3 times per week, so I wonder can we make stock for weekly solution, and keep it at 4'C in the fridge? Is CCl4+oil stable solution for few days?
Dear Milena,
I think you dont need keep it on fridge. You can make a stock solution and keep it on the bench
Dear all,
quick question, where did you get CCl4? do you use the powder or liquid version?
Thank you in advance!
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