Establishment of immortalized dental follicle cells for generating periodontal ligament in vivo
Establishment of immortalized dental follicle cells for generating periodontal ligament in vivo
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DOI：
10.1007/s00441-006-0257-6
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发表时间：
2007-02-01
影响因子：
3.6
通讯作者：
Teranaka, T.
中科院分区：
文献类型：
--
作者：
Yokoi, T.;Saito, M.;Teranaka, T.
牙囊是包围发育中的牙胚的间充质组织。在牙根形成过程中，牙周成分，即，牙骨质、牙周膜（PDL）和牙槽骨由牙囊祖细胞产生。在这里，我们报告的PDL祖细胞在小鼠牙囊细胞的存在。从小鼠切牙牙胚中获得细胞，并通过表达缺乏PDZ结构域结合基序的突变型人乳头状瘤病毒16型E6基因使细胞永生化。表达突变E6基因的MDFE 6-EGFP终止细胞（MDFE 6-EGFP superscript stop cells）具有延长的寿命，超过150个群体倍增（PD）。相比之下，正常的胰岛细胞不能增殖超过10 PD。MDFE 6-EGFP superscript终止细胞表达肌腱/韧带表型相关基因，如Scleraxis（Scx）、生长和分化因子-5、EphA 4、Six-1和I型胶原。另外，观察骨膜蛋白的表达。为了阐明MDFE 6-EGFP superscript终止细胞在体内的分化能力，将细胞移植到严重联合免疫缺陷小鼠中。在4周时，MDFE 6-EGFP superscript终止细胞移植物具有产生表达骨膜蛋白、Scx和XII型胶原以及I型胶原的纤维状组装的PDL样组织的能力。我们的研究结果表明，MDFE 6-EGFP superscript停止细胞可以作为PDL祖细胞，这些细胞可能是一个有用的研究工具，研究PDL的形成和开发再生疗法。
The dental follicle is a mesenchymal tissue that surrounds the developing tooth germ. During tooth root formation, periodontal components, viz., cementum, periodontal ligament (PDL), and alveolar bone, are created by dental follicle progenitors. Here, we report the presence of PDL progenitors in mouse dental follicle (MDF) cells. MDF cells were obtained from mouse incisor tooth germs and immortalized by the expression of a mutant human papilloma virus type 16 E6 gene lacking the PDZ-domain-binding motif. MDF cells expressing the mutant E6 gene (MDFE6-EGFP supercript stop cells) had an extended life span, beyond 150 population doublings (PD). In contrast, normal MDF cells failed to proliferate beyond 10 PD. MDFE6-EGFP supercript stop cells expressed tendon/ligament phenotype-related genes such as Scleraxis (Scx), growth and differentiation factor-5, EphA4, Six-1, and type I collagen. In addition, the expression of periostin was observed. To elucidate the differentiation capacity of MDFE6-EGFP supercript stop cells in vivo, the cells were transplanted into severe combined immunodeficiency mice. At 4 weeks, MDFE6-EGFP supercript stop cell transplants had the capacity to generate a PDL-like tissue that expressed periostin, Scx, and type XII collagen and the fibrillar assembly of type I collagen. Our findings suggest that MDFE6-EGFP supercript stop cells can act as PDL progenitors, and that these cells may be a useful research tool for studying PDL formation and for developing regeneration therapies.