The aim of this study was to extensively characterise natal dental pulp stem cells (nDPSC) and assess their efficiency to generate human induced pluripotent stem cells (hiPSC). A number of distinguishing features prompted us to choose nDPSC over normal adult DPSC, in that they differed in cell surface marker expression and initial doubling time. In addition, nDPSC expressed 17 out of 52 pluripotency genes we analysed, and the level of expression was comparable to human embryonic stem cells (hESC). Ours is the first group to report comprehensive characterization of nDPSC followed by directed reprogramming to a pluripotent stem cell state. nDPSC yielded hiPSC colonies upon transduction with Sendai virus expressing the pluripotency transcription factors POU5F1, SOX2, c-MYC and KLF4. nDPSC had higher reprogramming efficiency compared to human fibroblasts. nDPSC derived hiPSCs closely resembled hESC in terms of their morphology, expression of pluripotency markers and gene expression profiles. Furthermore, nDPSC derived hiPSCs differentiated into the three germ layers when cultured as embryoid bodies (EB) and by directed differentiation. Based on our findings, nDPSC present a unique marker expression profile compared with adult DPSC and possess higher reprogramming efficiency as compared with dermal fibroblasts thus proving to be more amenable for reprogramming.

Department of Biophysics Faculty of Medicine in Hradec Kralove Charles University Simkova 870 500 03 Hradec Kralove Czech Republic

Department of Dentistry Faculty Hospital in Hradec Kralove and Faculty of Medicine in Hradec Kralove Charles University Simkova 870 500 03 Hradec Kralove Czech Republic

Department of Histology and Embryology Faculty of Medicine in Hradec Kralove Charles University Simkova 870 500 03 Hradec Kralove Czech Republic

Department of Molecular Medicine Institute of Basic Medical Sciences University of Oslo 0317 Oslo Norway

Department of Oncology and Radiotherapy Faculty of Medicine in Hradec Kralove Charles University Simkova 870 500 03 Hradec Kralove Czech Republic

Department of Pediatric Research Oslo University Hospital 0424 Nydalen Norway

Department of Pharmacology Faculty of Medicine in Hradec Kralove Charles University Simkova 870 500 03 Hradec Kralove Czech Republic

Hybrid Technology Hub Centre of Excellence Institute of Basic Medical Sciences University of Oslo Blindern 0317 Oslo Norway

Institute of Immunology Oslo University Hospital Rikshospitalet PO Box 4950 Nydalen Oslo 0424 Norway

Norwegian Center for Stem Cell Research University of Oslo 0317 Oslo Norway

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