An In Vitro Method to Generate Astrocyte Scaffolds within Hydrogel Micro-columns
Transcrição
Take a culture dish containing a micro-column boat comprising hollow tubes of defined diameter.
The micro-columns, made of a hydrogel, are filled with a buffer to prevent drying.
Under a stereomicroscope, remove the buffer and add an ice-cold collagen solution, a component of the extracellular matrix.
Introduce a media along the dish's inner edge to maintain column hydration. Incubate under physiological conditions for the collagen to polymerize, forming a coating inside the micro-columns.
Add astrocytes inside the micro-columns and incubate to promote astrocyte adhesion to collagen.
Next, fill the dish with the medium and maintain it under physiological conditions.
Due to the micro-columns' narrow diameter, the astrocytes acquire a bipolar morphology with processes extending along the length.
Over time, the astrocytes self-assemble to form a three-dimensional scaffold containing dense bundles of astrocyte processes.
The hydrogel-encased astrocyte scaffold is ready for downstream assays.
