Kuroda Kengo, Kiyono Tohru, Eitsuka Takahiro, Isogai Hiroshi, Takahashi Koichi, Donai Kenichiro, Isogai Emiko, Fukuda Tomokazu
Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-amamiyamachi, Aoba-ku, Sendai, 981-8555, Japan.
J Cell Biochem. 2015 Feb;116(2):205-11. doi: 10.1002/jcb.24963.
Nonhuman primates are useful animal models for the study of human diseases. However, the number of established cell lines from nonhuman primates is quite limited compared with the number established from other experimental animals. The establishment of nonhuman primate cell lines would allow drug testing on those cell lines before moving experiments into primates. In this study, we established nonhuman primate primary cell lines by introducing the genes for CDK4R24C, cyclin D1, and hTERT. These cell lines proliferated more rapidly than primary cells and bypassed cellular senescence. Karyotype analysis showed that the chromosome patterns were intact in the immortalized cell lines. Furthermore, we showed that the expression of introduced genes could be precisely controlled through the Tet-Off system with the addition of doxycycline. The present study shows that introduction of the CDK4R24C, cyclin D1, and hTERT genes are effective methods of establishing nonhuman primate cell lines.
非人灵长类动物是研究人类疾病的有用动物模型。然而,与从其他实验动物建立的细胞系数量相比,从非人灵长类动物建立的细胞系数量相当有限。建立非人灵长类动物细胞系将允许在将实验转移到灵长类动物之前在这些细胞系上进行药物测试。在本研究中,我们通过引入CDK4R24C、细胞周期蛋白D1和hTERT的基因建立了非人灵长类动物原代细胞系。这些细胞系比原代细胞增殖更快,并且绕过了细胞衰老。核型分析表明,永生化细胞系中的染色体模式是完整的。此外,我们表明,通过添加强力霉素的Tet-Off系统可以精确控制导入基因的表达。本研究表明,引入CDK4R24C、细胞周期蛋白D1和hTERT基因是建立非人灵长类动物细胞系的有效方法。