Immortalization of cells derived from domestic dogs through expressing mutant cyclin-dependent kinase 4, cyclin D1, and telomerase reverse transcriptase

通过表达突变型细胞周期蛋白依赖性激酶 4、细胞周期蛋白 D1 和端粒酶逆转录酶,使源自家犬的细胞永生化。

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作者:Izzah Munirah,Taku Ozaki,Aya Sekine,Motoko Morimoto,Mayu Sugawara,Haruka Takada,Eriko Sugano,Hiroshi Tomita,Tohru Kiyono,Tomokazu Fukuda

Abstract

Dog is the first animal that was established as a close partner of human beings. Based on the vast genetic diversity and breeding, dogs exhibit unique genetic evolution and diversity from Chihuahua to St. Bernard. The safety tests of the pharmacological products also included domestic dogs as the test subjects. Although the safety confirmation test of chemicals for human use is important, the welfare of experimental animals requires special consideration. In this study, we cultured domestic dog-derived primary fibroblasts isolated from their muscle tissues. Furthermore, we successfully immortalized them through lentivirus-mediated gene transfer of mutant cyclin-dependent kinase 4 (CDK4), cyclin D1, and telomere reverse transcriptase (TERT). We further demonstrated that the established immortalized domestic dog-derived fibroblasts retained the characteristics of the original parental cells. These cells might act a suitable in vivo model system to replace the implication of animals for evaluating the potential toxicity of pharmacological chemicals. Supplementary information: The online version contains supplementary material available at 10.1007/s10616-021-00504-0.

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