Imaging Transgenic Beta-Cells in an Immunodeficient Mouse Challenged with Diabetic Splenocytes
Trascrizione
Begin with a restrained NOD immunodeficient mouse that lacks immune cells and is prone to diabetes.
This mouse is pre-injected with transgenic pancreatic beta-cells that produce insulin and express luciferase enzymes.
Take a syringe containing pre-treated splenocytes derived from a diabetic mouse. The splenocytes include autoreactive T cells with the ability to recognize the insulin peptides.
Administer these splenocytes into the mouse's tail vein.
The splenocytes circulate and reach the pancreas, where the autoreactive T cells recognize the insulin peptides on the beta cells and become activated.
These activated cells release cytotoxic molecules, initiating beta-cell destruction via autoreactivity.
Next, inject a luciferase substrate into the peritoneum of the mouse. The substrate diffuses in this region and enters the beta cells.
Within the cells, the substrate interacts with the luciferase enzymes, producing luminescence.
Using a bioluminescent imaging system, capture images at regular intervals
A gradual decrease in luminescence indicates the destruction of transgenic beta-cells due to autoreactivity.
