I know it must be subcutaneous but where? and if any one have picture or video it will be ok
First of all, a specific type of mouse has to be utilized, such as female nude mouse or female SCID mouse, since these types of mice have suppressed immune systems. These mice should be injected either into the nipples or into their flanks on one side or both sides. Cells to be injected should be mixed with about equal volume of Matrigel prior to their inoculation. One should also be careful to determine the time needed for xenografts to appear depending on a number of tumor cells used for injection, since some tumors can regressed, while others would progress.
Thanks for you very much and i want to ask:
1- is their any way to use the normal mice in this experiment?
2- how can i suppress immune systems of normal ones?
3- how can i prepare Matrigel?
Thanks a lot
We use a variety of breast cell lines in SCID female mice. You can either perform subcutaneous xenografts or orthotopic injections, into the site of the breast. If you anesthetize the mice, gently place the needle tip, under the nipple area (we normally use the the 3rd nipple down, to prevent difficulty in mobility, when tumour is growing) and inject into the fat pad of the mammary tissue. This is a good way to achieve metastasis studies too. Any further quieries please feel free to contact me.
MAtrigel preparation is 50:50 with cells. Matrigel must be stored in fridge 2 days prior to use, to allow the gel to liquify, makes it easier to use. Keep all syringes, tips and pippettes on ice or keep in fridge prior to use.
For subcutaneous inject on the right flank of the mouse (right side of the abdomen, just under the skin). Be careful not to scratch underlying tissues as this can lead to an attached tumour, which can not be used for statistical analysis.
We do breast cancer xenografts commercially (http://altogenlabs.com/xenograft-models/breast-cancer-xenograft/) and our standard procedure is to inject into the hind flank with matrigel. Immunodeficient mice are a necessity, as an active immune system can be problematic for tumor growth. Cyclosporin treatment can be used to suppress some immune responses, but for preclinical studies it is worth investing into immunodeficient models.
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