I would like to reduce the numbers of BM cells through irradiation and reconstitute with new BM cells, or not. The goal would be to study whether peripheral immune cell precursors are able to regenerate the hematopoetic system without BM-precursors.
Sounds like you might want to look at shielding. If you're working with mice, make sure the radiation hits the femur and tibia bones, where most of the bone-marrow lives. Shield the peripheral organs with a lead cutout. And then use some kind of dosimetry (photo film?) to make sure you got what you wanted.
Total Body Irradiation (TBI) with 11Gy (Cesium ) is the best to clear of bone marrow. We Use C57BL6 wild type mice for this purpose as they have proven radiosenitivity. after that you should use sterile environment for them and give them betryl as an adjuvant in drinking water for at least 3-5 days to prevent any bacterial infections.
The best is you give TBI at least 24Hrs before you planned for Bone marrow transplantation through retro orbital route.
best of luck
Hi Rodney, you mean 'irradiation'?
As said above, shielding is important - protect the brain if you can, and give whole body irradiation of 2 x 7.5 Gy, with 24 h in between. I understand that you don't actually want functional bone marrow in your mice?
Cheers,
Thomas
Thank all for the answers... I will try out the shielding technique as suggested.
You may wish to check the 1960's literature from the Paterson Labs, Manchester, England, particularly the papers of LG Lajtha (a founder of the field of stem cell kinetics) , N. Testa, R Schofield (originator of stem cell niche hypothesis), D. Porteous, etc. They did lots of TBI and BM rescue studies, measured the amount of marrow in each bone (in mice), etc.
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