Isolation and Culture of Primary Schwann Cells from a Human Dermis
Transcription
Take pieces of isolated human dermis containing various cell types, including fibroblasts, Schwann cells, and immune cells, embedded within a collagen matrix.
Mince the tissue into small fragments.
Add collagenase, a proteolytic enzyme, to digest the tissue's collagen fibers, loosening the cells.
Mechanically dissociate the tissue, obtaining a cell suspension.
Pass the cells through a cell strainer, removing the cell aggregates and tissue debris.
Add media containing serum to the cell suspension to stop the collagenase activity.
Centrifuge and discard the supernatant containing collagenase.
Resuspend the cells in media and plate them in a poly-L-lysine-coated culture flask.
Incubate for the cells to adhere to the coated surface.
Remove the media containing non-adherent cells. Wash with buffer.
Incubate with an antimitotic agent to eliminate rapidly dividing cells, like fibroblasts.
Remove media. Wash with buffer to remove any remaining antimitotic agents.
Add Schwann cell-specific culture media and incubate, facilitating the growth of Schwann cells.
