Kawano Eisuke, Toriumi Taku, Iguchi Shinya, Suzuki Daigo, Sato Shuichi, Honda Masaki
Division of Applied Oral Sciences, Nihon University Graduate School of Dentistry.
Department of Anatomy, Nihon University School of Dentistry.
Biomed Res. 2017;38(2):135-147. doi: 10.2220/biomedres.38.135.
We previously generated induced pluripotent stem (iPS) cells from human dental pulp cells of deciduous teeth. Neural crest cells (NCCs) play a vital role in the development of the oral and maxillofacial region. Therefore, NCCs represent a cell source for bone, cartilage, and tooth-related tissue engineering. In this study, we examined whether iPS cells are capable of differentiating into NCCs through modification of the human embryonic stem cell protocol. First, iPS cells were dissociated into single cells and then reaggregated in low-cell-adhesion plates with neural induction medium for 8 days in suspension culture to form neurospheres. The neurospheres were transferred to fibronectin-coated dishes and formed rosette structures. The migrated cells from the rosettes abundantly expressed NCC markers, as evidenced by real-time polymerase chain reaction, immunofluorescence, and flow cytometric analysis. Furthermore, the migrated cells exhibited the ability to differentiate into neural crest lineage cells in vitro. They also exhibited tissue-forming potential in vivo, differentiating into bone and cartilage. Collectively, the migrated cells had similar characteristics to those of NCCs. These results suggest that human dental pulp cell-derived iPS cells are capable of differentiating into NCCs. Therefore, iPS cell-derived NCCs represent cell sources for bone and cartilage tissue engineering.
我们之前从乳牙的人牙髓细胞中诱导生成了诱导多能干细胞（iPS细胞）。神经嵴细胞（NCCs）在口腔颌面部发育中起着至关重要的作用。因此，NCCs是骨、软骨及牙齿相关组织工程的细胞来源。在本研究中，我们通过修改人类胚胎干细胞方案，研究了iPS细胞是否能够分化为NCCs。首先，将iPS细胞解离为单细胞，然后在低细胞粘附板中与神经诱导培养基重聚，在悬浮培养中培养8天以形成神经球。将神经球转移到纤连蛋白包被的培养皿中并形成玫瑰花结结构。来自玫瑰花结的迁移细胞大量表达NCC标志物，实时聚合酶链反应、免疫荧光和流式细胞术分析证明了这一点。此外，迁移细胞在体外表现出分化为神经嵴谱系细胞的能力。它们在体内也表现出组织形成潜力，可分化为骨和软骨。总的来说，迁移细胞具有与NCCs相似的特征。这些结果表明，人牙髓细胞来源的iPS细胞能够分化为NCCs。因此，iPS细胞来源的NCCs是骨和软骨组织工程的细胞来源。
