(PO39) Differentiation of IPSC for Potential Cell-Based Regeneration

Abstract: This study addresses the challenge of dental pulp regeneration by investigating the potential of induced pluripotent stem cells (iPSCs) derived from human dental pulp stem cells (hDPSCs). Reprogramming of hDPSCs into iPSCs was achieved using the Epi5^TM Episomal iPSC Reprogramming Kit. The pluripotency of the reprogrammed iPSCs was confirmed through differentiation into endoderm, mesoderm, and ectoderm, validated by immunofluorescence staining. Characterization of the iPSCs was conducted using real-time quantitative polymerase chain reaction (RT-qPCR), western blot, and immunofluorescence staining. Embryoid bodies (EBs) were generated from iPSCs for subsequent differentiation into endothelial and epithelial cells, with further characterization using RT-qPCR, western blot, and immunofluorescence staining. Successful iPSC colonies were formed around day 41, signifying readiness for differentiation. The pluripotency of the reprogrammed iPSCs was confirmed through differentiation into the three germ layers, validated by immunofluorescent staining. RT-qPCR and western blot analysis confirmed the successful reprogramming of hDPSCs to iPSCs. iPSC-derived endothelial and epithelial cells exhibited distinct characteristics typical of their respective lineages. The angiogenic potential was confirmed through a tube formation assay. This study presents systematic protocols for the transformation of hDPSCs into iPSCs, followed by differentiation into endothelial and epithelial cells. iPSCs can be applied in dental fields, especially in tooth structure formation.