利用PiggyBac转座子系统构建NIS-EGFP-Fluc三报告基因人胚胎干细胞系
Generation of a NIS-EGFP-Fluc triple-reporter human embryonic stem cell line by PiggyBac transposon system.
作者信息
Huang Pufeng, Tian Chuanhuizi, Chen Yanyan, Zhou Meng, Ni Jie, Niu Mengda, Liu Xujie, Peng Zhiping
机构信息
Department of Radiological Medicine and Oncology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, China; Shenzhen Key Laboratory of Cardiovascular Disease, Fuwai Hospital Chinese Academy of Medical Sciences, Shenzhen, Shenzhen 518057, Guangdong Province, China.
Department of Radiological Medicine and Oncology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
出版信息
Stem Cell Res. 2022 Dec;65:102968. doi: 10.1016/j.scr.2022.102968. Epub 2022 Nov 14.
One major challenge in stem cell therapy is to longitudinally track cell fate after cells transplantation. Molecular Imaging approaches enabling noninvasive long-term monitoring the transplanted cells are imperative for assessment of the safety and efficiency. Here, we used PiggyBac technology to insert triple reporter genes: NIS, EGFP and Firefly luciferase into a human embryonic stem cell line (hESCs, H9) and obtained a reporter hESCs line (NIS-EGFP-Fluc H9). The triple-reporters allows the transplanted NIS-EGFP-Fluc H9 cells and their derivates to be fluorescence, bioluminescence and even PET/SPECT imaged. This triple-reporter hESCs line provides a valuable imaging platform for cell-based therapeutics clinical translation.
干细胞治疗中的一个主要挑战是在细胞移植后纵向追踪细胞命运。能够对移植细胞进行无创长期监测的分子成像方法对于评估安全性和有效性至关重要。在此，我们使用PiggyBac技术将三重报告基因：NIS、EGFP和萤火虫荧光素酶插入人胚胎干细胞系（hESCs，H9）中，获得了一个报告基因hESCs系（NIS-EGFP-Fluc H9）。这三重报告基因使得移植的NIS-EGFP-Fluc H9细胞及其衍生物能够进行荧光、生物发光甚至PET/SPECT成像。这个三重报告基因hESCs系为基于细胞的治疗临床转化提供了一个有价值的成像平台。
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