Bovine and porcine fibroblasts can be immortalized with intact karyotype by the expression of mutant cyclin dependent kinase 4, cyclin D, and telomerase
Bovine and porcine fibroblasts can be immortalized with intact karyotype by the expression of mutant cyclin dependent kinase 4, cyclin D, and telomerase
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DOI：
10.1016/j.jbiotec.2014.02.017
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发表时间：
2014-04-20
影响因子：
4.1
通讯作者：
Fukuda, Tomokazu
中科院分区：
文献类型：
--
作者：
Donai, Kenichiro;Kiyono, Tohru;Fukuda, Tomokazu
牛和猪是经济上最重要的牲畜。尽管它们的重要性，从这些物种培养的细胞，这是有用的生理分析，在细胞库是相当有限的。细胞系数量有限的原因之一是它们的建立困难。为了克服细胞系建立的局限性，我们试图通过多种细胞周期调节因子（突变细胞周期蛋白依赖性激酶4、细胞周期蛋白D和端粒酶逆转录酶）的转导使牛和猪成纤维细胞永生化。转导后的细胞继续表现出稳定的增殖速率，没有出现细胞衰老。此外，细胞周期分析表明，这些外源基因的诱导增强了细胞周期的周转，特别是在G1-S期。此外，我们建立的细胞系维持正常的二倍体核型在98-100%。我们的研究表明，绕过p16/ rb介导的细胞阻滞和端粒酶活性的激活，可以有效地建立永活的牛和猪源成纤维细胞。建立细胞系的高效率表明，细胞周期调控因子网络，特别是p16/ rb相关的细胞周期阻滞，在人类、牛和猪的进化过程中得到了保存。(C) 2014 Elsevier B.V.版权所有
Cattle and pigs comprise the most economically important livestock. Despite their importance, cultured cells from these species, which are useful for physiological analyses, are quite limited in cell banks. One of the reasons for the limited number of cell lines is the difficulty in their establishment. To overcome limitations in cell-line establishment, we attempted to immortalize bovine and porcine fibroblasts by transduction of multiple cell cycle regulators (mutant cyclin dependent kinase 4, cyclin D and telomerase reverse transcriptase). The transduced cells continued to display a stable proliferation rate and did not show cellular senescence. Furthermore, cell cycle assays showed that induction of these exogenous genes enhanced turnover of the cell cycle, especially at the G1-S phase. Furthermore, our established cell lines maintained normal diploid karyotypes at 98-100%. Our study demonstrated that bypassing p16/Rb-mediated cell arrest and activation of telomerase activity enabled efficient establishment of immortalized bovine- and porcine-derived fibroblasts. The high efficiency of establishing cell lines suggests that the networks of cell cycle regulators, especially p16/Rb-associated cell cycle arrest, have been conserved during evolution of humans, cattle, and pigs. (C) 2014 Elsevier B.V. All rights reserved.