ct26 tumor does not grow in mice - please help! (Dec/04/2008 )
Hi there,
I have problems in growing tumor after injecting CT26 cells in C57BL/6 mice. Briefly I am counting the cells when they have ~60-70 confluency and I am spinning the cells and then resuspending them in ice cold PBS and then I mixed them(~3 millions cells for each injection) with Matrigel in a total volume of 500ul and, using a 26 G needle I inject this sc in flank. But the tumors are not growing. What can be the problem: the cells, I am destroying the cells when inoculate them, my technique? Any suggestions?
Thank you very much!
I have problems in growing tumor after injecting CT26 cells in C57BL/6 mice. Briefly I am counting the cells when they have ~60-70 confluency and I am spinning the cells and then resuspending them in ice cold PBS and then I mixed them(~3 millions cells for each injection) with Matrigel in a total volume of 500ul and, using a 26 G needle I inject this sc in flank. But the tumors are not growing. What can be the problem: the cells, I am destroying the cells when inoculate them, my technique? Any suggestions?
Thank you very much!
Hello,
My first thought is whether the C57BL/6 mouse is the appropriate syngeneic mouse strain. If your CT26 cell line is the mouse colon adenocarcinoma line I'm thinking of, then to my knowledge BALB/C mice or BALB/C x DBA/2 F1 (CD2F1) mice are the appropriate strains. My impression is that tumor take can also be affected by: (1) the duration/harshness of the trypsinization procedure used to harvest cells (I'm kind of paranoid about this, so I try to limit trypsin exposure as much as possible); (2) shearing of cells (we use 25 G needles for sc injections, but I wouldn't think 26G would make a difference); (3) passage number (I believe some cells can experience decrements in in vivo tumorigenicity with time in culture); (4) time between harvest and injection into mice.
I hope this is of some help. Good luck!
SK
Thanks SK,
I will try these things. I am also planning to buy new cells because I do not exactly the history of the present stock. I hope this will help.
I have read articles where they used C57Bl/6 mice and they got good ct26 tumors so until now I have not thought about this. I will do some search to be sure.
Thanks again!
Dan
Hi SamOH
I did some reseacrch and I thing you are right. Actually I was wrong when I said that other people were using this combination (ct26 cell and C57Bl/6 mice). Nobody use them! So this can be my probplem. Can you explain me more about this incompatibility because I couldn't found details about this online.
Thank you very much!
Dan
I did some reseacrch and I thing you are right. Actually I was wrong when I said that other people were using this combination (ct26 cell and C57Bl/6 mice). Nobody use them! So this can be my probplem. Can you explain me more about this incompatibility because I couldn't found details about this online.
Thank you very much!
Dan
Hi Dan,
I hope it really is as simple as changing the strain of mouse. I do not know the specifics off-hand, but it is akin to an immunological rejection of a transplant. The tumor cells have antigens that the host recognizes as foreign.
Glad I could help - good luck.
SamOH