Induction of neural crest cells from human dental pulp-derived induced pluripotent stem cells
Induction of neural crest cells from human dental pulp-derived induced pluripotent stem cells
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DOI：
10.2220/biomedres.38.135
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发表时间：
2017-01-01
影响因子：
1.2
通讯作者：
Honda, Masaki
中科院分区：
文献类型：
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作者：
Kawano, Eisuke;Toriumi, Taku;Honda, Masaki
我们以前产生诱导多能干细胞（iPS）从人乳牙牙髓细胞。神经嵴细胞（NCCs）在口腔颌面部的发育过程中起着至关重要的作用。因此，NCC代表了骨、软骨和牙齿相关组织工程的细胞来源。在这项研究中，我们研究了iPS细胞是否能够通过修改人胚胎干细胞方案分化为NCC。首先，将iPS细胞解离成单细胞，然后在具有神经诱导培养基的低细胞粘附板中悬浮培养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。实时聚合酶链反应、免疫荧光和流式细胞术分析表明，从罗氏细胞中迁移的细胞大量表达NCC标记。此外，迁移的细胞在体外表现出向神经嵴谱系细胞分化的能力。它们在体内也表现出组织形成的潜力，分化成骨和软骨。总的来说，迁移的细胞具有与NCC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NCC。因此，iPS细胞衍生的NCC代表骨和软骨组织工程的细胞来源。
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.