Decellularized Matrix Derived from Neural Differentiation of Embryonic Stem Cells Enhances the Neurogenic Potential of Dental Follicle Stem Cells

Introduction: Dental follicle stem cells (DFSCs) possess neurogenic potential because they originate from the embryonic neural:crest. This study investigated whether neural differentiation of DFSCs can be enhanced by culture on decellularized matrix substrata (NSC-DECM) derived from neurogenesis of human embryonic stem cells (hESCs). Methods: The hESCs were differentiated into neural stem cells (NSCs), and NSC-DECM was extracted from confluent monolayers of NSCs through treatment with deionized water. DFSCs seeded on NSC-DECM, Geltrex, and tissue culture polystyrene (TCPS) were subjected to neural induction during a period of 21 days. Expression of early/intermediate (Musashi1, PAX6, NSE, and beta 111-tubulin) and mature/late (NGN2, NeuN, NFM, and MASH1) neural markers by DFSCs was analyzed at the 7-, 14-, and 21-day time points with quantitative real-time polymerase chain reaction. Immunocytochemistry for detection of beta 111-tubulin, PAX6, and NGN2 expression by DFSCs on day 7 of neural induction was also carried out. Results: Quantitative RT-PCR showed that expression of PAX6, Musashi1, beta llI-tubulin, NSE, NGN2, and NFM by DFSCs was enhanced on NSCDECM versus either the Geltrex or TCPS groups. Immunocytochemistry showed that DFSCs in the NSC-DECM group displayed more intense staining for beta lll-tubulin, PAX6, and NGN2 expression, together with more neurite outgrowths and elongated morphology, as compared with either Geltrex or TCPS. Conclusions: DECM derived from neurogenesis of hESCs can enhance the neurogenic potential of DFSCs.