Functional Odontoblastic-Like Cells Derived from Human iPSCs

• Published in: Journal of dental research 2018-01, Vol.97 (1), p.77-83
• Main Authors: Xie, H., Dubey, N., Shim, W., Ramachandra, C.J.A., Min, K.S., Cao, T., Rosa, V.
• Format: Article
• Language: English
• Subjects: Alkaline phosphatase; Animals; Biological activity; Biomedical materials; Cell differentiation; Cell Differentiation - drug effects; Cell self-renewal; Dental pulp; Dental Pulp - drug effects; Dental Pulp - physiology; Dentin; Dentistry; Deoxyribonucleic acid; DNA; Electroporation; Female; Gene expression; Humans; Immunodeficiency; Induced Pluripotent Stem Cells - physiology; KLF4 protein; Kruppel-Like Transcription Factors - pharmacology; Mice; Mice, SCID; Mineralization; Myc protein; Oct-4 protein; Octamer Transcription Factor-3 - pharmacology; Odontoblasts; Odontoblasts - metabolism; Odontoblasts - physiology; Phosphatase; Pluripotency; Polylactic acid; Proto-Oncogene Proteins c-myc - pharmacology; RNA-Binding Proteins - pharmacology; SOXB1 Transcription Factors - pharmacology; Stem cell transplantation; Stem cells; Tissue engineering; Tissue Scaffolds
• ISSN: 0022-0345; 1544-0591
• DOI: 10.1177/0022034517730026

The induced pluripotent stem cells (iPSCs) have an intrinsic capability for indefinite self-renewal and large-scale expansion and can differentiate into all types of cells. Here, we tested the potential of iPSCs from dental pulp stem cells (DPSCs) to differentiate into functional odontoblasts. DPSCs were reprogrammed into iPSCs via electroporation of reprogramming factors OCT-4, SOX2, KLF4, LIN28, and L-MYC. The iPSCs presented overexpression of the reprogramming genes and high protein expressions of alkaline phosphatase, OCT4, and TRA-1-60 in vitro and generated tissues from 3 germ layers in vivo. Dentin discs with poly-L-lactic acid scaffolds containing iPSCs were implanted subcutaneously into immunodeficient mice. After 28 d from implantation, the iPSCs generated a pulp-like tissue with the presence of tubular dentin in vivo. The differentiation potential after long-term expansion was assessed in vitro. iPSCs and DPSCs of passages 4 and 14 were treated with either odontogenic medium or extract of bioactive cement for 28 d. Regardless of the passage tested, iPSCs expressed putative markers of odontoblastic differentiation and kept the same mineralization potential, while DPSC P14 failed to do the same. Analysis of these data collectively demonstrates that human iPSCs can be a source to derive human odontoblasts for dental pulp research and test bioactivity of materials.