Immortalized cementoblasts and periodontal ligament cells in culture
Immortalized cementoblasts and periodontal ligament cells in culture
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DOI：
10.1016/s8756-3282(99)00096-4
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发表时间：
1999-07-01
期刊：
影响因子：
4.1
通讯作者：
Somerman, MJ
中科院分区：
文献类型：
--
作者：
D'Errico, JA;Ouyang, H;Somerman, MJ
牙骨质是一种附着在牙根表面的矿化组织，在牙周发育过程中是形成功能性牙周韧带附着物所必需的。此外，在疾病后的组织再生过程中，骨质被认为在修复过程中起关键作用。研究工作旨在了解牙周发育和再生的机制，特别是牙根骨质的形成，一直受到无法分离和培养参与牙骨质产生的细胞，即成骨水泥细胞的阻碍。使用传统的成骨细胞分离技术，从牙根表面的细胞中建立了永活的，异质的成骨水泥细胞/牙周韧带细胞（CM/PDL）群体：1) CD-1小鼠，使用SV40使细胞永生化；2)H-2K(b)tsA58“永生化”小鼠，其中含有永生化转基因的细胞被移除并培养。CM/PDL群体来源于生长中的牙根表面粘附的组织，而粘附在周围牙槽骨上的组织被明确排除在群体之外。对永生化CM/PDL细胞进行了表征，以确保其表型反映了先前在原位和原代非永生化培养中所证明的表型。与骨/骨质相关的蛋白质/ mrna，已知由根衬骨水泥母细胞表达，而不是由原位PDL细胞表达，例如骨涎蛋白、骨桥蛋白和骨钙素，在永生化群体中的细胞中表达。此外，CM/PDL细胞在体外以精氨酸-甘氨酸-天门氨酸（RGD）依赖的方式附着在骨涎蛋白上，促进矿物结节的形成，并表现出PTH/ pthrp介导的cAMP反应。这些含有CR I和PDL细胞的永生化异质群体为研究参与骨质形成的细胞提供了独特的机会，并增强了我们对牙周组织发育、维持和再生的控制机制的认识。(C) 1999年Elsevier Science Inc.版权所有。
Cementum, a mineralized tissue lining the surface of the tooth root, is required for formation of a functional periodontal ligament attachment during development. Additionally during regeneration of tissues after disease, cementum is thought to play a critical role in the reparative process. Research efforts aimed toward understanding mechanisms involved in periodontal development and regeneration, and in particular the formation of root cementum, have been hampered by an inability to isolate and culture cells involved in cementum production, i.e., cementoblasts, Using classical techniques for osteoblast isolation, immortalized, heterogeneous cementoblast/periodontal ligament cell (CM/PDL) populations were established from cells lining the tooth root surface of: 1) CD-1 mice, where cells were immortalized using SV40, or 2) H-2K(b)tsA58 "immorto" mice, where cells containing an immortalizing transgene were removed and cultured. CM/PDL populations were derived from tissues adherent to developing tooth root surfaces, while tissues adherent to the surrounding alveolar bone were specifically excluded from the population. Immortalized CM/PDL cells were characterized to ensure their phenotype reflected that previously demonstrated in situ and in primary, nonimmortalized cultures, Proteins/mRNAs associated with bone/cementum and known to be expressed by root lining cementoblasts, but not by PDL cells, in situ, e.g., bone sialoprotein, osteopontin, and osteocalcin, were expressed by cells within the immortalized populations. Furthermore, CM/PDL cells, in vitro, attached to bone sialoprotein in an arginine-glycine-aspartic acid (RGD)-dependent manner, promoted mineral nodule formation and exhibited a PTH/PTHrP-mediated cAMP response, These immortalized heterogeneous populations, containing both CR I and PDL cells, provide a unique opportunity to study cells involved in cementogenesis and to enhance our knowledge of the mechanisms controlling development, maintenance, and regeneration of periodontal tissues. (C) 1999 by Elsevier Science Inc. rill rights reserved.