Preparation of a Micropatterned Substrate to Study Schwann Cell Phenotypes
Transcription
Take a silicon wafer with micropatterns of the desired geometry to grow cells in a defined shape.
Pour a polymer mixture on it and allow it to solidify.
Cut a square stamp of the polymer.
Coat the stamp's patterned surface with a protein layer, which facilitates cell attachment.
Using an air stream, remove unbounded proteins.
Take a polymer-coated coverslip.
Place the patterned side of the stamp on it.
Press the stamp to transfer the pattern. Remove the stamp.
Transfer the coverslip into a well containing a surfactant to coat the bare surfaces. This increases surface hydrophobicity, preventing unwanted cell adhesion.
Remove the surfactant and wash with a phosphate buffer.
Add the Schwann cells and allow them to attach to the micropattern.
Wash with a buffer to remove unattached cells.
Add culture medium and incubate.
As the cells grow along the surface, they take the micropattern shape, enabling a study of Schwann cell phenotype.
