Effect of Poly-L-lysine Coating on Retinoic Acid-Loaded PLGA Microspheres in the Differentiation of Carcinoma Stem Cells into Neural Cells

Abstract

In this study, PLGA microspheres were prepared using a water-in-oil-in-water emulsion/solvent evaporation technique. Some microspheres were coated with poly-L-lysine (an extracellular matrix (ECM) component), and then pluripotent P19 embryonic carcinoma cells were seeded on them. P19 cells attached onto the PLGA microspheres; subsequently, by adding retinoic acid (RA) to cell culture medium as a neurogenic inducer (RA was released from the microspheres), the cells differentiated into neural cells. Size and morphology of PLGA microspheres was characterized by scanning electron microscopy (SEM). Neurogenic differentiation was studied by immunofluorescent staining, real-time polymerase chain reaction (RT-PCR), and light microscopy. Histological assay showed that more cells attached onto microspheres coated with poly-L-lysine than the uncoated group. Immunofluoresent staining and RT-PCR analysis for β-Tubulin, Nestin and Pax6 genes indicated differentiation of P19 cells into neural cells on both coated and uncoated microspheres. It was found that a high surface area of microspheres improves cell attachment and expansion, which was significantly increased in those coated with poly-L-lysine. Finally, these results highlight the versatility of these sample scaffolds as a model system for nerve tissue engineering.