I am developing a cancer mouse model. I injected a pancreatic cancer cell with two variant WT and KO(some gene) in nude mice subcutaneously. After 4-5 days all mice died in KO batch but all mice are still alive in WT group. What could be the possible explanation for that?
We do pancreatic cancer xenografts commercially (http://altogenlabs.com/xenograft-models/pancreatic-xenograft-models/) and we standardize our procedures based on the cell line. It is possible that the KO version differs so much from the WT one that the protocols for both may have to differ to optimize tumor growth. We inject in the hind leg with matrigel, but I have never seen circumstances such as yours. Let us know how the followup xenografts go and that might give us more clues to give advice.
probably you have to reduce num of cells in ko group for injection.
Hi nikhil, unfortunately i won't trust on that experiment because of the short period in wich all Ko animals died. you should reduce the amount of cells inyected , also perform the inyection the same day in all animals.
check for infection signals in all animals
Thank you for your feedback guys but the problem is i already injected low number of cells(1.5 million). Experiment will not be feasible if i inject lower than that and i performed the same day everything plus i also check the contamination for the ko cells. Anyway i will repeat the experiment to make sure the problem is real. Meanwhile if you have any comments or suggestion please let me know.
Thanks a lot
First What is de gene you were working with? Second: didi you perform an autopsy ? Third why do not try again?
Thank you for your feedback Prof. I am already planning for the second try andt the gene i am working with is ERO1A. Actually i did not get a chance to perform an autopsy because when i went for follow up animal staff already removed the dead mice and i just saw only one mice also dead (what i saw was mice shrinked a lot).
where the WT and KO mice in separated cages? (one for KO and one for WT)? Could it be a cage problem? Or there were multiple cages?
By the way, The best is always to have animals representative of both groups in the same cage.
Did the animals injected with KO cells develop any tumors?
Where did you implant them exactly? Maybe the location might cause this problem. That would be nice to open them and see.
We do pancreatic cancer xenografts commercially (http://altogenlabs.com/xenograft-models/pancreatic-xenograft-models/) and we standardize our procedures based on the cell line. It is possible that the KO version differs so much from the WT one that the protocols for both may have to differ to optimize tumor growth. We inject in the hind leg with matrigel, but I have never seen circumstances such as yours. Let us know how the followup xenografts go and that might give us more clues to give advice.
I would suggest the KO cells were infected. Do you test them for e.g. mycoplasma prior to injection?
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