Differentiation of Mouse Stem Cells into Neural Cells on PLGA Microspheres Scaffold

The cellular therapy and nerve tissue engineering will probably become a major therapeutic strategy for promoting axonal growth through injured area in central nervous system and peripheral nervous system in the coming years. The stem cell carrier scaffolds in nerve tissue engineering resulted in strong survival of cells and suitable differentiation into neural cells, so this pathway should be created a favorable environment for axon regeneration. Poly lactic-co-glycolic acid (PLGA) has been widely used for manufacturing three dimentional scaffolds for tissue engineering. The pluripotent nature and proliferative capacity of embryonic carcinoma cells such as P19 also makes them an attractive cell source for tissue engineering. This study was initiated to evaluate potential of biodegradable PLGA microspheres for P19-derived neurons for neural tissue engineering and axon regeneration. The PLGA microspheres were prepared by using solvent evaporation, water in oil in water, technique. The water phase was polyvinyl alcohol (PVA) solution and the oil phase was PLGA solution. Retinoic acid (RA) was added to bacterial dishes as a differentiation factor inducer. P19 cells were attached to the PLGAmicrospheres and differentiated into neural cells on them. PLGA microspheres were characterized for size and surface morphology by scanning electron microscopy. The in vitro experimental studies were performed via immunoflouresent staining, scanning electron microscopy (SEM), RT-PCR, and histology. The photomicrograph and histology staining show the surrounded microspheres by P19 cells. The SEM results demonstrated the attachment and axon formation. Immunoflouresent staining and RT-PCR analysis for MapII, -Tubulin, Nestin and Pax6 indicated the differentiation of P19 cells into neural cells. This report shows that high surface area also allows rapid cell expansion and increases cell attachment on PLGA microspheres, so each microsphere contains high cell density that resulted in survival of transplantation into the straitum of host animals, therefore, PLGA microspheres can help the differentiation of P19 cells into neural cells.