I am planning to do orthotopic injection of breast cancer cells (MDA-MB-231 and 4T1 cells) into the mammary fat pad of mice to observe cancer metastasis and tumor growth. In lots of references I see people used growth factor reduced (GFR) matrigel and mixed it with the cells. My question is what is the purpose of using GFR matrigel. Will it makes a different when using matrigel containing growth factor?
If the tumor cells grow well subcutaneously, they should grow well in the foot-pad without the need of any enhancer.
The secret for the successful transplantation is the foot fad is to inject extremely small volumes of cell suspensions. In other words, the foot pad is an extremely small space to inject anything. You will feel resistance immediately during the injection. So, your cell suspension must be highly concentrated, so that if only 10 or 20 ul go inside the pad, there are enough cells to grow.
The leg needs to be amputated after the tumor has reached a 0.5 cm or so. Then wait for the metastases to become visible to the naked eye or at the microscope.
Hope this helps.
Frank Arguello
atavisticchemotherapy.com
GRF-Matrigel was reported to stimulate proliferation more than matrigel. But how GF are reduced isn't clear in most manufacturer pamphlets.
I have injected 4T1 cells into the mammary fat pad of mouse without any materigel and it worked absolutely fine.
I have also developed the tumor only by subcutaneous injection of 4T1 cells close to the mammary fat pad area in the mouse thigh and the tumor grew absolutely fine. 4T1 is very aggressive in nature and mostly does not require any matrigel.
You should not need to use Matrigel when injecting into the mammary fat pad - as others have said, 4T1 will grow almost anywhere without the additional ECM support. Depending on your experiment, matrigel can potentially confound your analysis. It can be difficult to determine that you have implanted into the mammary fat pad rather than sub-cutaneously near the fat pad, since the cells grow equally well in either site.
Dear all respondents Can I grow MDA-MB-231 cells with out matrigel by injecting into the mammary fat pad?
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