Cultivation and imaging of astrocytes on protein-coated Fluorescent topographies constructed from aligned PLLA electrospun fibers

Electrospun poly- L -lactic acid (PLLA) fibers are presently explored as tissue engineering platforms for regeneration of the central nervous system. In particular, aligned, electrospun fibers are capable of directing astrocyte cellular extension and migration. The precise mechanisms by which aligned, electrospun substrates alter glial cell behavior are poorly understood. Therefore, there is a need for designing and refining electrospun fiber platforms and developing novel approaches for studying astrocytic behavior and physiology on aligned substrates. Here, we describe and discuss methods for (1) fabrication of fluorescent PLLA microfibers by electrospinning, (2) coating PLLA fibers with different extracellular matrix (ECM) proteins to facilitate attachment of astroglial cells, (3) isolation of primary astrocytes and plating them onto PLLA fibers, and (4) imaging the interactions between PLLA fibers and astrocytes to better understand the ability of fibers to enable astrocyte extension and migration.