title: Isolation and Characterization of Human Adult Epithelial Stem Cells from the Periodontal Ligament.
abstract: We report a novel method for the isolation of adult human epithelial stem cells (hEpiSCs) from the epithelial component of the periodontal ligament-the human epithelial cell rests of Malassez (hERM). hEpiSC-rich integrin-α6(+ve) hERM cells derived by fluorometry can be clonally expanded, can grow organoids, and express the markers of pluripotency (OCT4, NANOG, SOX2), polycomb protein RING1B, and the hEpiSC supermarker LGR5. They maintain the growth profile of their originating hERM in vitro. Subcutaneous cotransplantation with mesenchymal stem cells from the dental pulp on poly-l-lactic acid scaffolds in nude mice gave rise to perfect heterotopic ossicles in vivo with ultrastructure of dentin, enamel, cementum, and bone. These remarkable fully mineralized ossicles underscore the importance of epithelial-mesenchymal crosstalk in tissue regeneration using human progenitor stem cells, which may have already committed to lineage despite maintaining hallmarks of pluripotency. In addition, we report the clonal expansion and isolation of human LGR5(+ve) cells from the hERM in xeno-free culture conditions. The genetic profile of LGR5(+ve) cells includes both markers of pluripotency and genes important for secretory epithelial and dental epithelial cell differentiation, giving us a first insight into periodontal ligament-derived hEpiSCs.
authors.affiliation: Laboratory of Tooth Organogenesis and Regeneration, Department of Orthodontics and Pediatric Medicine, School of Dentistry; Center for Organogenesis, School of Medicine; Center for Computational Medicine and Bioinformatics; University of Michigan, Ann Arbor, MI, USA Laboratory for Oral, Head, and Neck Cancer Metastasis, Kresge Hearing Research Institute, Department of Otolaryngology and Comprehensive Cancer Center, Medical School, University of Michigan, Ann Arbor, MI, USA The Cancer Cure, Ann Arbor, MI, USA.
authors.forename: M
authors.initials: M
authors.lastname: Athanassiou-Papaefthymiou
authors.affiliation: Laboratory of Tooth Organogenesis and Regeneration, Department of Orthodontics and Pediatric Medicine, School of Dentistry; Center for Organogenesis, School of Medicine; Center for Computational Medicine and Bioinformatics; University of Michigan, Ann Arbor, MI, USA hellene@umich.edu.
authors.forename: P
authors.initials: P
authors.lastname: Papagerakis
authors.affiliation: Laboratory for Oral, Head, and Neck Cancer Metastasis, Kresge Hearing Research Institute, Department of Otolaryngology and Comprehensive Cancer Center, Medical School, University of Michigan, Ann Arbor, MI, USA Department of Periodontics and Oral Medicine, School of Dentistry, University of Michigan, Ann Arbor, MI, USA.
authors.forename: S
authors.initials: S
authors.lastname: Papagerakis
mesh_terms: D053687:Adult Stem Cells / Q000502:physiology*; D000818:Animals; D004847:Epithelial Cells / Q000166:cytology* / Q000502:physiology; D018398:Homeodomain Proteins / Q000502:physiology; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000502:physiology; D010513:Periodontal Ligament / Q000166:cytology* / Q000502:physiology; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology; D063150:Polycomb Repressive Complex 1 / Q000502:physiology; D043562:Receptors, G-Protein-Coupled / Q000502:physiology; D055748:SOXB1 Transcription Factors / Q000502:physiology; D033581:Stem Cell Transplantation; D054457:Tissue Scaffolds; D014074:Tooth Calcification / Q000502:physiology
chemical_list: D018398:Homeodomain Proteins; C116884:LGR5 protein, human; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D043562:Receptors, G-Protein-Coupled; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D063150:Polycomb Repressive Complex 1; C489841:RNF2 protein, human
keywords: Lgr5; cell rests of Mallassez; cementum; dental pulp stem cells; enamel; integrin-α6