Department of Cardiology, Leiden University Medical Center, Leiden, the Netherlands.
Department of Cardiothoracic Surgery, Leiden University Medical Center, Leiden, the Netherlands.
Nat Biomed Eng. 2022 Apr;6(4):389-402. doi: 10.1038/s41551-021-00827-5. Epub 2022 Jan 6.
The lack of a scalable and robust source of well-differentiated human atrial myocytes constrains the development of in vitro models of atrial fibrillation (AF). Here we show that fully functional atrial myocytes can be generated and expanded one-quadrillion-fold via a conditional cell-immortalization method relying on lentiviral vectors and the doxycycline-controlled expression of a recombinant viral oncogene in human foetal atrial myocytes, and that the immortalized cells can be used to generate in vitro models of AF. The method generated 15 monoclonal cell lines with molecular, cellular and electrophysiological properties resembling those of primary atrial myocytes. Multicellular in vitro models of AF generated using the immortalized atrial myocytes displayed fibrillatory activity (with activation frequencies of 6-8 Hz, consistent with the clinical manifestation of AF), which could be terminated by the administration of clinically approved antiarrhythmic drugs. The conditional cell-immortalization method could be used to generate functional cell lines from other human parenchymal cells, for the development of in vitro models of human disease.
缺乏可规模化且稳健的高分化人类心房肌细胞来源限制了心房颤动 (AF) 的体外模型的发展。在这里，我们展示了通过一种依赖于慢病毒载体的条件细胞永生化方法，以及在人胎心房肌细胞中使用强力霉素控制表达重组病毒癌基因，可以生成和扩增 10 的 24 次方倍的功能齐全的心房肌细胞，并且可以使用永生化细胞来生成 AF 的体外模型。该方法生成了 15 个具有类似于原代心房肌细胞的分子、细胞和电生理特性的单克隆细胞系。使用永生化心房肌细胞生成的多细胞体外 AF 模型显示出纤维颤动活性（激活频率为 6-8 Hz，与 AF 的临床表现一致），可通过给予临床批准的抗心律失常药物来终止。该条件细胞永生化方法可用于从其他人类实质细胞生成功能性细胞系，以开发人类疾病的体外模型。
