I read a study (link below) where they cultured cells and re-suspended them in a buffer before injection. I am curious what buffers are safe for this? PBS? aMEM? What do you think?
I think PBS should be ok.Its a composition of sodium and potassium based and will not interfere with the safety of the host.Maybe normal saline could be a better bet than PBS as it is a solution of simple NaCl.
PBS is pretty safe. Also Ringer's, Tyrode, Hank's balanced salt solution, etc.
(you forgot the link)
I have used PBS, DPBS, HBSS and EBSS in vivo in the past with no complications. In addition it is not uncommon to use a % of BSA in PBS as a buffer for cells in an injection formulation.
Just a reminder that anything you inject into an animal (experimental, therapeutic, cells), to include the buffer that it is prepared in, should be approved by the IACUC prior to doing so. There are quite a few considerations that should be made with the exact buffer (most likely normal saline or PBS), pH (solutions with higher or lower pH will cause tissue necrosis if injected IM or SC), route, etc...As of 2011 with the new Guide requirements, anything adminstrered other than pharmaceutical grade compounds also requires IACUC approval with scientific justifcation (not including parsimonious reasons).
We used sterile PBS (Vehicle group), For treatment group = Cell preparation in melting medium (alpha MEM+HSA) and finally cells suspended in sterile PBS were infused.
In medical field, for most of the intravenous drugs saline or sometimes ringer lactate solution are used. But for intramuscular injection most of the times distilled water is used. As Mr. Steingrimur Stefansson mentioned other solution also can be used based on the requiremient.
We have used Tris buffer at pH 7.5 in mice injections and it worked fine (that buffer was required by the protein we injected). However, as mentioned above, PBS should work fine too. Make sure that you filter sterilize the solution to inject. In our case, we had to run our protein through an endotoxin removal column too.
Tris buffer + EDTA is recommended. EDTA acts as a chelator in biologic milieu.
Have not seen problems with either Ringer's or PBS. Both are isotonic and have been used in vivo but the question is how does a researcher choose among the two? Is it just plane random choice?
Hi Ellie, if you are going to inject cells into animals, the best way is to use tissue culture grade media, DMEM, EMEM, whatever.
These are sterile media that have minimal endotoxin and other pyrogens in them.
You just have to be careful about using sterile techniques to prep. the cells for injection.
Please could you just clarify whether the 0.1 Molar (i.e 10x) PBS is isotonic
Judit Marshillach Lopez, have you guys published that? I have my protein in Tris 20mM and would like to ip inject it into mice. I could dilute it even a bit in PBS, but I want to make sure there won't be any unspecific effect.
Thanks!
We SQ and IP inject proteins in 20mM Tris Buffer into mice/rats quite frequently, and have seen no effects.
This is a very subjective discussion. The mode of injection is also a prime criteria to be discussed and differentiated together with different buffers used. PBS seems to do not much i.p but intra tracheal, intra dermal etc..there are local effects. It depends on what one calls no effect.
Though I do not use intra dermal PBS injection in my experiments, I am curious to know experience from other people for the same method, same buffer without any cells or chemicals.
I highly recommend a paper by S.C. Gad et al (2006), entitled, Nonclinical Vehicle Use in Studies by Multiple Routes in Multiple Species. This paper was published in the Int. J. of Tox. 2006, 25: 499.
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