Purifying Capillaries From a Porcine Brain
Transkript
Take a porcine brain.
Remove the meninges, the protective membrane covering the brain, to prevent meningeal cell contamination.
Scrape the grey matter, a region rich in capillaries, and collect the tissue in a culture dish containing media.
Pass the grey matter through a syringe without a needle to fragment the tissue.
Transfer the fragments to a homogenizer and homogenize to disintegrate the tissue further, releasing the brain capillaries.
Pass the homogenate through a filter with a specific pore size to retain the capillaries, allowing other brain cells to flow through.
Place the filter in a solution containing enzymes and DNase.
The enzymes detach the adherent cells, while DNase degrades contaminating DNA, facilitating the isolation of pure capillaries.
Wash the capillaries from the filter.
Add media containing serum proteins to inactivate the enzymes.
Transfer the capillaries to a tube, centrifuge, and remove the supernatant. Resuspend the pure capillaries in fresh media for further analysis.
