I have fibroblasts in culture from 4 week old mice. I am trying the 3T3 protocol on them, and am currently at the tenth passage. However, as the 3T3 protocol was developed for MEFs I am wondering if it might work on my cells, or if they will simply die at some point.
I used a differentiation protocol that worked both for the 3T3 cells and stem cells from adult rat mammary gland. It worked on both cell types. I'm sending you the paper.
Thank you for your answer! However what I have are not 3T3 cells, but primary fibroblasts from transgenic mice that I am trying to immortalize using the 3T3 protocol. What I am wondering is if the 3T3 protocol only works on embryonic cells, or if it will also work on my cells from 4 week old mice.
"fibroblast" could be a little bit not precise, because I think there is not a cell that actually is only a fibroblast. In my opinion there is no need for "fibroblasts" in a living being. What it will be their job? It is very likely that the stuff call in scientific papers are in reality totipotent or multi-potent stem cells, waiting just for the right signal to differentiate into an useful cell type(epithelial, osteoblast, osteoclast, vascular and so on). I do not think that there will be a problem with age, I got my cells from post-pubescent rats and I could passage them many times, always having the same behavior, without even try to "immortalize" them. 3T3 that are sold actually came from a mush of mice embryos, so who knows really what are their capability? So that is probably the reason why they behave as stem cells.
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