Fluorescent topographies constructed from aligned PLLA electrospun fibers

Electrospun poly- L -lactic acid (PLLA) fi bers are presently explored as tissue engineering platforms for regeneration of the central nervous system. In particular, aligned, electrospun fi bers 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 refi ning electrospun fi ber platforms and developing novel approaches for studying astrocytic behavior and physiology on aligned substrates. Here, we describe and discuss methods for (1) fabrication of fl uorescent PLLA microfi bers by electrospinning, (2) coating PLLA fi bers with different extracellular matrix (ECM) proteins to facilitate attachment of astroglial cells, (3) isolation of primary astrocytes and plating them onto PLLA fi bers, and (4) imaging the interactions between PLLA fi bers and astrocytes to better understand the ability of fi bers to enable astrocyte extension and migration.