Hello everyone! i have two queries.
1) I intend to develop the tumor subcutaneously in BALB/c mice using HT29 cells. I will not use Matrigel along with the cells. how much minimum number of cells are enough to develop the tumor? and is there any upper limit too?
2). why the age of mice is important, what if my mice are above 3 months old?
Hi Asrhad, i have little experience in colorectal cancer but this is what I think.
Regarding the first question, as Nicole said, you must try several dilutions of your HT29 cells in order to stablish the efficacy of the engraftment, tumor volume, speed of growth etc. in our subcutaneous experiment (pancreatic cells) we determine that 1x 10^6 or 2x 10^6 were the correct amount of cells to generate very consistent tumors, we also try lower amount of cells and we did not get tumors in most of the mice, so you have to perform this pilot experiment or refer to the bibliography of your specific HT29 cells.
Regarding the 2 question, age is very important for BALB/C nude mice because despite those mice have a serious impairment in T-cells there are other very important lineages of immune cells that can appear as the mice get older as the macrophages m1 / m2, NK, MDSC and others, and that immune compartment can interfere with your results, so I would recommend to start the xenograft subcutaneous experiment at 6- 7 weeks of age.
Best.
Thank you Nicole and Carlos for your input, i have injected cells with serial dilution. Have a good weekend)
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