Immortalization of cementoblast progenitor cells with Bmi-1 and TERT
Immortalization of cementoblast progenitor cells with Bmi-1 and TERT
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DOI：
10.1359/jbmr.041006
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发表时间：
2005-01-01
影响因子：
6.2
通讯作者：
Teranaka, T
中科院分区：
文献类型：
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作者：
Saito, M;Handa, K;Teranaka, T
从用Bmi-1和hTERT永生化的牙囊细胞中成功分离出命名为BCPb 8的成牙骨质细胞祖细胞系。BCPb 8在植入免疫缺陷小鼠后显示出分化为成牙骨质细胞的潜力。BCPb 8是第一个建立的成牙骨质细胞祖细胞系，为研究牙骨质形成提供了一个有用的模型。在牙根发育过程中，存在于牙囊中的祖细胞被认为在牙周组分（牙骨质、牙周韧带和牙槽骨）的形成中起核心作用。然而，对这些祖细胞的生物学知之甚少。以前，我们观察到培养的牛牙囊细胞（BDFC）含有推定的成牙骨质细胞祖细胞。为了进一步分析这些细胞的生物学特性，我们试图通过表达多梳组蛋白Bmi-1和人端粒酶逆转录酶（hTERT.Materials and Methods）从永生化BDFC中分离成牙骨质细胞祖细胞。单细胞克隆建立从永生化BDFC，并分化成成牙骨质细胞进行评估植入到immunodeficiency.Results和结论：BDFCs表达Bmi-1和hTERT表现出延长的寿命-90人口doubles超过正常BDFCs-和仍然含有细胞的潜力，分化成牙骨质细胞植入到免疫缺陷小鼠。从这些细胞中，我们建立了一个克隆细胞系，命名为BCPb 8，形成牙骨质样组织，是反应性的抗牙骨质特异性单克隆抗体3G 9和表达mRNA的骨唾液酸蛋白，骨钙素，骨桥蛋白，和I型胶原植入。因此，通过使用Bmi-1和hTERT，我们成功地使来自BDFC的成牙骨质细胞祖细胞永生化而不影响分化潜能。BCPb 8细胞系是第一个成牙骨质细胞祖细胞的永生化克隆细胞系，不仅可以用于研究牙骨质形成，而且还可以用于开发牙周炎患者的再生治疗。
A cementoblast progenitor cell line designated BCPb8 was successfully isolated from dental follicle cells immortalized with Bmi-1 and hTERT. BCPb8 showed the potential to differentiate into cemento-blasts on implantation into immunodeficient mice. BCPb8 was confirmed to be the first established cementoblast progenitor cell line and will provide a useful model for investigating cementogenesis.Introduction: The dental follicle is the mesenchymal tissue surrounding the developing tooth germ. During tooth root development, progenitor cells present in the dental follicle are believed to play a central role in the formation of periodontal components (cementum, periodontal ligament, and alveolar bone). However, little more is known about the biology of these progenitors. Previously, we observed that cultured bovine dental follicle cells (BDFCs) contained putative cementoblast progenitors. To further analyze the biology of these cells, we attempted to isolate cementoblast progenitors from immortalized BDFC through expression of the polycomb group protein, Bmi-1, and human telomerase reverse transcriptase (hTERT).Materials and Methods: BDFCs were transduced with replication-deficient retroviruses carrying human Bmi-1(LXSN-Bmi-1), and hTERT (LXSH-hTERT) for immortalization. Single cell clones were established from immortalized BDFC, and differentiation into cementoblasts was assessed by implantation into immunodeficient mice.Results and Conclusion: BDFCs expressing Bmi-1 and hTERT showed an extended life span-90 population doublings more than normal BDFCs-and still contained cells with the potential to differentiate into cementoblasts on implantation into immunodeficient mice. From these cells, we established a clonal cell line, designated BCPb8, which formed cementum-like tissue that was reactive to the anti-cementum-specific monoclonal antibody 3G9 and expressed mRNA for bone sialoprotein, osteocalcin, osteopontin, and type I collagen on implantation. Thus, by using Bmi-1 and hTERT, we succeeded in immortalizing cementoblast progenitor cells from BDFC without affecting differentiation potential. The BCPb8 cell line is the first immortalized clonal cell line of cementoblast progenitors and could be a useful tool not only to study cementogenesis but also to develop regeneration therapy for patients with periodontitis.