A Method for the High Salt Treatment of Dendritic Cells and their Adoptive Transfer into Mice
Transcript
Begin with a multi-well plate containing a suspension of dendritic cells or DCs.
To mimic a hypertensive condition, add a culture medium containing a high concentration of sodium ions and incubate.
During incubation, sodium ions enter the cell, increasing the intracellular sodium concentration.
This triggers the exchange of intracellular sodium ions with extracellular calcium ions.
Increased internal calcium levels activate pathways that facilitate the generation of reactive aldehydes, which bind to the cellular proteins. This activates the DCs, which display these modified proteins on their surface.
Post-incubation, collect the cells in a tube and centrifuge.
Remove the supernatant. Add fresh buffer to resuspend the cells.
Draw the cell suspension into a syringe.
Take an anesthetized mouse and inject the cell suspension retroorbitally which is an area behind the eye socket.
This process of injecting activated immune cells is called adoptive cell transfer.
