Li Xiangfen, Wang Liu, Su Qin, Ye Ling, Zhou Xuedong, Song Dongzhe, Huang Dingming
State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China.
Department of Endodontics, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China.
Stem Cells Int. 2020 Feb 15;2020:4534128. doi: 10.1155/2020/4534128. eCollection 2020.
Human dental pulp cells (HDPCs) play a vital role in dentin formation and reparative dentinogenesis, which indicated their potential application in regenerative medicine. However, HDPCs, which can only be obtained from scarce human pulp tissues, also have a limited lifespan , and stem cells usually lose their original characteristics over a large number of passages. To overcome these challenges, we successfully immortalized human dental pulp cells using the piggyBac system which was employed to efficiently overexpress the SV40 T-Ag, and we then comprehensively described the cell biological behavior. The immortalized human dental pulp cells (iHDPCs) acquired long-term proliferative activity and expressed most HDPC markers. The iHDPCs maintained multiple differentiation potential and could be induced to differentiate into chondrogenic, osteogenic, and adipogenic cells . We also proved that the iHDPCs gained a stronger ability to migrate than the primary cells, while apoptosis was inhibited. Furthermore, highly proliferative iHDPCs displayed no oncogenicity when subcutaneously implanted into athymic nude mice. Finally, iHDPCs exhibited odontogenic differentiation ability and secreted dentin sialophosphoprotein (DSPP) when combined with a beta-tricalcium phosphate scaffold and bone morphogenetic protein-2 (BMP2) . Conclusively, the established iHDPCs are a valuable resource for mechanistic study of dental pulp cell differentiation and dental pulp injury repair, as well as for applications in tooth regeneration.
人牙髓细胞（HDPCs）在牙本质形成和修复性牙本质生成中起着至关重要的作用，这表明它们在再生医学中具有潜在的应用价值。然而，HDPCs只能从稀缺的人牙髓组织中获取，其寿命也有限，并且干细胞通常会在大量传代后失去其原始特性。为了克服这些挑战，我们使用piggyBac系统成功地使人牙髓细胞永生化，该系统用于高效过表达SV40 T-Ag，然后我们全面描述了细胞生物学行为。永生化人牙髓细胞（iHDPCs）获得了长期增殖活性并表达了大多数HDPC标志物。iHDPCs保持了多种分化潜能，可被诱导分化为软骨细胞、成骨细胞和脂肪细胞。我们还证明，iHDPCs比原代细胞具有更强的迁移能力，同时细胞凋亡受到抑制。此外，高度增殖的iHDPCs皮下植入无胸腺裸鼠时未显示出致瘤性。最后，iHDPCs与β-磷酸三钙支架和骨形态发生蛋白-2（BMP2）联合时表现出牙源性分化能力并分泌牙本质涎磷蛋白（DSPP）。总之，所建立的iHDPCs是牙髓细胞分化和牙髓损伤修复机制研究以及牙齿再生应用的宝贵资源。
