Isolation and Culturing of Cells from Mouse Embryo Dorsolateral Telencephalons
Transcription
Take a mouse embryo in a buffer and separate the skull to isolate the brain.
Remove the meninges, the membranous layer covering the brain.
Dissect the brain to separate the hemispheres.
Isolate the dorsolateral telencephalons from the hemispheres and transfer them into a tube.
The dorsolateral telencephalons are rich in neurons and neural progenitor cells.
Centrifuge and remove the supernatant.
Add an enzyme solution. The enzymes break down the tissue matrix, loosening the cells.
Introduce a proliferation medium containing serum. The serum proteins inhibit enzyme activity.
Use a pipette to mechanically dissociate the tissue, forming a cell suspension.
Centrifuge and remove the supernatant containing inactivated enzymes and tissue debris.
Resuspend the cells in the proliferation medium.
Transfer the cells onto a cell culture substrate-coated microplate.
Incubate for the cells to adhere to the substrate. The medium facilitates progenitor cell proliferation.
