I have tried 2x106 and 5x106 cells of EL-4 to inoculate in C57 mice，but when I use 2x106 the tumor could not form and when I use 5x106 the tumor will disappear at day 7 or 8.what's wrong with the cells?
Sometimes is needed adding corticosteroids in the médium to make a graft. Do you try it?
No experience with this particular cell line but know it was chemically induced and has been around for many decades. The description of your transplantation experience suggests it's probably become antigenic. If you are much more successful in NOD/SCID mice or even nude mice, it would be a strong indicator of immune rejection in your C57 mice. Could also whole body irradiate the C57 mice to supress it's immune system to get an answer, as well (perhaps 4-5 Gy - check literature).
It depends.
For a subcutaneous tumor, with 100,000 cells you will have a tumor palpable in 5-7 days. If not, your cells maight be infected with a virus, thus, are more immunogenic
Hi Yuan
I have experience with EL4 and their more immunogenic version E.G7 (transfected with OVA) and I have found that these cells readily form tumors in C57 mice (8 - 10 wks of age) at challenges of 1x106 - 1x107 subQ. I think you have mycoplasma contaminating your cell line - have you checked for mycoplasma? There are kits available for this. If you find your cells are contaminated I would get a new batch of EL4 cells (ideally from ATCC).
If your cells are actually mycoplasma negative then, barring some other form of contamination or cross contamination with another cell line, I would think that some technical aspect of the tumor challenge is to blame. Make sure you challenge with cells that are in log-phase of growth ( not overgrown) and that they are >90% viable. Also challenge with cells that are suspended in PBS or media that contains no foetal calf serum since that could act as an immunogen.
good luck
Sean
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