I'm planning on injecting some melanoma and hepatoma cell lines mixed with Matrigel into the mice subcutaneously. However, I'm not sure what the pros and cons of diluting Matrigel with HBSS, RPMI, PBS, or other solutions are.
You can use serum free media or PBS. I generally use 1:1 mixture of matrigel. If you prefer to use lower amount, then you can dilute the matrigel beforehand with either serum free media or PBS. Make sure to thaw the marigel on ice. Also, once thawed on ice, make sure the matrigel is homogeneous by slowly mixing before you add that to the cell suspension. I usually use the cell suspension (2X) to dilute matrigel. For eg. if I am injecting 100 ul of cell suspecsion of certain density, I prepare the cell suspension which is twice that density, take 500 ul of that in a fresh eppendorf and add 500 ul of undiluted matrigel to get a 1:1 mixture. Mix slowly without generating bubbles. You can draw 100 ul (or your preferred volume) for each injection. Good luck.
Thanks Swati!
Instinctively I would think serum free RPMI is more superior, when compared to PBS, for injection. It probably gives a bit more nutrients for the cell to start growing.
However, I'm not sure whether RPMI will encourage certain tumour cells to grow and result in a tumour that is genetically different to a tumour grew in PBS in mice.
Hi Simon,
As you are injecting only a few micro liters, either one is OK. Any medium should work just fine (serum free), as after injection, the growth will be driven by mostly in vivo factors such as immunity and how aggressive the cell line is. Of course, matrigel is ussful to hold the cells together and prevent them leaking out of the injection site. When you inject, make sure to hold the needle inside for a few minutes. The mouse's body temp allow the matrigen with cells to solidify quite quickly. Good luck!. I use growth factor reduced matrigel for this procedure. The cencer cells also produce their own growth factors which drive their growth as well. Good luck!
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