Generating Neural Progenitors from Embryonic Stem Cells in Serum-Free Monolayer Cultures
Transcription
Begin with a culture of mouse embryonic stem cells.
Wash the cells with a buffer to remove media and cell debris.
Add an enzyme cocktail to detach cells from the flask surface.
Add media to stop the enzymatic activity and collect the cells in a tube. Centrifuge and discard the supernatant containing enzymes.
Resuspend the cells in serum-free neuronal culture media.
Seed the cells onto gelatin-coated multi-well plate wells.
Incubate. The embryonic stem cells adhere to the gelatin-coated surface. Media constituents, including nutrients, growth factors, and hormones, facilitate cell proliferation and form a monolayer.
At this cell density, the embryonic stem cells produce and release optimal levels of growth factors and cytokines.
These molecules trigger intracellular signaling pathways, facilitating the differentiation of embryonic stem cells into neural progenitors.
