Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-Sen University, Guangzhou, Guangdong 510080, PR China.
Biomaterials. 2013 Feb;34(6):1701-11. doi: 10.1016/j.biomaterials.2012.11.018. Epub 2012 Dec 4.
Pluripotent stem cells, including embryonic stem (ES) and induced pluripotent stem (iPS) cells, serve as unlimited resources for cell replacement therapy and tissue engineering because such cells are capable of extensive proliferation in vitro and can give rise to lineages that represent any of the three embryonic germ layers. However, in the context of the in vivo behavior of cell transplants, key challenges need to be addressed and essential strategies should be developed before stem cells can be used in clinical practice. In the present study, we modified mouse ES/iPS cells to contain a suicide gene, deltaTK or CodA, under the transcriptional control of the EF1α or Nanog promoter. The suicide gene was introduced via lentivirus transduction without interfering with their self-renewal and pluripotency characteristics. We found that EF1α promoter-controlled deltaTK/CodA expression efficiently eliminated pluripotent stem cells and their derivatives both in vitro and in vivo. When the suicide gene was under the control of the Nanog promoter, tumor-initiating undifferentiated pluripotent stem cells were selectively ablated in vitro after prodrug treatment. These results indicate that modification of pluripotent stem cells with a suicide gene prior to transplantation offers a safe manner by which wayward stem cells, and their progeny, can be controlled in vivo. Our approach will render the clinical application of human pluripotent stem cells increasingly possible.
多能干细胞，包括胚胎干细胞（ES）和诱导多能干细胞（iPS），可作为细胞替代治疗和组织工程的无限资源，因为这些细胞能够在体外进行广泛增殖，并能产生代表三个胚胎胚层的谱系。然而，在细胞移植的体内行为背景下，需要解决关键挑战，并在干细胞能够在临床实践中应用之前制定必要的策略。在本研究中，我们通过慢病毒转导将自杀基因 deltaTK 或 CodA 修饰到小鼠 ES/iPS 细胞中，使其受 EF1α 或 Nanog 启动子的转录控制。自杀基因的引入不干扰其自我更新和多能性特征。我们发现，EF1α 启动子控制的 deltaTK/CodA 表达可有效消除体外和体内的多能干细胞及其衍生物。当自杀基因受 Nanog 启动子控制时，在药物前体处理后，体外肿瘤起始的未分化多能干细胞可被选择性清除。这些结果表明，在移植前对多能干细胞进行自杀基因修饰可提供一种安全的方法，以控制体内异常的干细胞及其后代。我们的方法将使人类多能干细胞的临床应用变得越来越可行。
