Hello,
I have generated DO11.10 / Rag1 KO mice with deletion of gene of my interest. So it is double gene KO and TCR transgenic.
I tried to isolate naive T cells from the mouse using FACS with control as heterozygote of my gene of interest, and found out that CD4+CD62L+DO11.10+ population is only ~1% of total splenocyte in both KO and heterozygote groups.
In absolute cell number, there were less than 10^6 naive T cells per single spleen of 6wk old mouse.
I have no experience in TCR transgenic + Rag deficient mice so I wonder if the low cell number is normal or there is something wrong with my mice.
If you have done experiments with such mice, please share your idea with me.
Thank you.
Hello Wonyong,
those mice do have very low splenocyte counts. May I ask, what where your total splenocyte numbers? As far as I know DO11 should have around 12% CD4+ of total splenocytes, so your absolute T cell number would fit with what I am used to have in total splenocyte numbers from those guys. But I do not quite understand how you come to only 1% in the FACS since you would have to have enormous amounts of splenocytes... just remember: DO11 basically only have naive T cells unless they have seen OVA.
Thank you Nicole for your answer. I haven't done counting total splenocyte yet, but spleens from those mice were really small, about 1/4 of age matching wt mouse. 1% came out from RBC-lysed total splenocytes. I thought most of splenocytes would be CD4+KJ1-26+ naive T cells since there would be no B cells or CD8+ T cells due to Rag1 deficiency, and did not do negative selection. Apprently I was wrong. 99% of them I have no idea what they are, but if the absolute number of T cells seems right, it would not matter much. Thanks again!
TCR transgenic lymphocytes compete for available endogenous ligands to survive. So, this is normal, if you see lymphopenia. This may be different in mice with another genetical background.
Thank you Daniel and Dmitry for your input. I guess my strain is unique in low cell number. It could be a reason why so little publications using it maybe.
- Badges
- Science topic
- Similar topics
- Medicine
- Disease
- Disease Models
- Animal Models
- Rodent Models
- Mouse Models