i am new in cancer immunology. could anyone help me with understanding following experiments?
"CBA/Ca mice received a total of two, four, or six i.p. injections at biweekly
intervals of MC57G-Vo, MC57G-24, MC57G-26 (single cell suspensions
of 107 cells in 200 ul), or PBS (200 ul). Four days after the last injection,
spleens were harvested. Lymphocytes were used as responders (5 105
cells/well) in an MLR with irradiated C57BL/6 splenocytes (3000 rad,
1.25 105 cells/well). Cultures were performed in triplicate wells using
96-well flat-bottom plates in 200 l of IMDM complete medium with
2-ME (50 M). Proliferation was assessed by adding [3H]thymidine (1
Ci/well) during the last 8–12 h of a 4-day culture. "
why the slpenocyte need to be irradiated?
Hi,
the spleenicytes are irradiated for them to not proliferate and so they will not incorporate the thimidyne. Like that your thymidine counts will only come from the proliferation of the responder lymphocytes.
cheers.
