Efficient immortalization of human dental pulp stem cells with expression of cell cycle regulators with the intact chromosomal condition
Efficient immortalization of human dental pulp stem cells with expression of cell cycle regulators with the intact chromosomal condition
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DOI：
10.1371/journal.pone.0229996
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发表时间：
2020-03-02
期刊：
影响因子：
3.7
通讯作者：
Fukuda, Tomokazu
中科院分区：
文献类型：
--
作者：
Orimoto, Ai;Kyakumoto, Seiko;Fukuda, Tomokazu
近年来的临床研究表明，自体牙髓干细胞动员移植是一种安全有效的牙髓再生治疗方法。然而，需要解决一些局限性，例如在移植前对分离的个体牙髓细胞产品进行安全性和质量控制测试的高成本。因此，更有效的体外培养人牙髓干细胞可能有助于为牙髓再生治疗提供低成本和高可靠性的测试。在这项研究中，我们建立了一个新的永生化牙髓干细胞系，共表达突变的细胞周期蛋白依赖性激酶4（R24 C），细胞周期蛋白D1，端粒酶逆转录酶（TERT）。建立的细胞系保持了其原有的二倍体染色体和干细胞特性，并表现出增强的增殖率。此外，我们发现永生化的人牙髓干细胞在适当的培养条件下，即使细胞增殖加速，仍然保持其成骨和成脂分化能力。综上所述，我们建立的细胞系可以作为牙髓再生治疗的有用体外工具，并且可以有助于细胞处理的再现性和简便性，从而节省与安全性和质量控制测试相关的时间和成本。
Clinical studies have recently demonstrated that autologous transplantation of mobilized dental pulp stem cells is a safe and efficacious potential therapy for pulp regeneration. However, some limitations need to be addressed, such as the high cost of the safety and quality control tests for isolated individual dental pulp cell products before transplantation. Therefore, more efficient in vitro culturing of human dental pulp stem cells might be useful for providing low cost and high reliability testing for pulp regeneration therapy. In this study, we established a novel immortalized dental pulp stem cell line by co-expressing a mutant cyclin-dependent kinase 4 (CDK4(R24C)), Cyclin D1, and telomerase reverse transcriptase (TERT). The established cell line maintained its original diploid chromosomes and stemness characteristics and exhibited an enhanced proliferation rate. In addition, we showed the immortalized human dental pulp stem cells still keeps their osteogenic and adipogenic differentiation abilities under appropriate culture conditions even though the cell proliferation was accelerated. Taken together, our established cell lines could serve as a useful in vitro tool for pulp regeneration therapy, and can contribute to reproducibility and ease of cell handling, thereby saving time and costs associated with safety and quality control tests.