Culture splenocytes
After the mice were completely anesthetized by sevoflurane, the mice
were killed by cervical dislocation. Then place the mouse in a bottle
filled with alcohol for 5 minutes to sterilize, and operate in a sterile
biological operation table: use a sterile instrument to perform
laparotomy to remove the spleen tissue of the mouse and place it into a
sterile mill previously added with sterile PBS Grind gently in the
grinding dish to completely separate the spleen cells, then add
erythrocyte lysate and let stand for 10 minutes, then stop with sterile
PBS, centrifuge to remove the lower layer of cells. After filtering out
the non-cellular components with a filter, count 1×106cells into a sterile culture dish, add Leukocyte Activation Cocktail 1ul
and incubate for 6 hours at 37 ° C with 5% CO2.