Athanassiou-Papaefthymiou M, Papagerakis P, Papagerakis S
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.
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
J Dent Res. 2015 Nov;94(11):1591-600. doi: 10.1177/0022034515606401. Epub 2015 Sep 21.
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.
我们报告了一种从牙周膜的上皮成分——人马拉瑟上皮剩余（hERM）中分离成人人类上皮干细胞（hEpiSCs）的新方法。通过荧光测定法获得的富含hEpiSC的整合素-α6（阳性）hERM细胞可以进行克隆扩增，能够生长类器官，并表达多能性标志物（OCT4、NANOG、SOX2）、多梳蛋白RING1B以及hEpiSC超级标志物LGR5。它们在体外维持其起源的hERM的生长特征。在裸鼠体内，将其与来自牙髓的间充质干细胞在聚左旋乳酸支架上进行皮下共移植，产生了具有牙本质、釉质、牙骨质和骨超微结构的完美异位小骨。这些显著的完全矿化的小骨强调了上皮-间充质相互作用在使用人类祖干细胞进行组织再生中的重要性，这些细胞尽管保持着多能性特征，但可能已经确定了细胞谱系。此外，我们报告了在无血清培养条件下从hERM中克隆扩增和分离人LGR5（阳性）细胞。LGR5（阳性）细胞的基因图谱既包括多能性标志物，也包括对分泌性上皮细胞和牙上皮细胞分化重要的基因，这使我们对牙周膜来源的hEpiSCs有了初步了解。
