Hausburg Frauke, Müller Paula, Voronina Natalia, Steinhoff Gustav, David Robert
Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center; Department Life, Light and Matter of the Interdisciplinary Faculty, Rostock University.
Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, Rostock University Medical Center.
J Vis Exp. 2018 Jun 18(136):57474. doi: 10.3791/57474.
While CD133 hematopoietic stem cells (SCs) have been proven to provide high potential in the field of regenerative medicine, their low retention rates after injection into injured tissues as well as the observed massive cell death rates lead to very restricted therapeutic effects. To overcome these limitations, we sought to establish a non-viral based protocol for suitable cell engineering prior to their administration. The modification of human CD133 expressing SCs using microRNA (miR) loaded magnetic polyplexes was addressed with respect to uptake efficiency and safety as well as the targeting potential of the cells. Relying on our protocol, we can achieve high miR uptake rates of 80-90% while the CD133 stem cell properties remain unaffected. Moreover, these modified cells offer the option of magnetic targeting. We describe here a safe and highly efficient procedure for the modification of CD133 SCs. We expect this approach to provide a standard technology for optimization of therapeutic stem cell effects and for monitoring of the administered cell product via magnetic resonance imaging (MRI).
虽然CD133造血干细胞已被证明在再生医学领域具有巨大潜力，但它们注射到受损组织后的低保留率以及观察到的大量细胞死亡率导致治疗效果非常有限。为了克服这些限制，我们试图在给药前建立一种基于非病毒的合适细胞工程方案。针对人CD133表达干细胞的摄取效率、安全性以及细胞的靶向潜力，研究了使用负载微小RNA（miR）的磁性多聚体对其进行修饰。依靠我们的方案，我们可以实现80%-90%的高miR摄取率，而CD133干细胞特性不受影响。此外，这些修饰后的细胞提供了磁靶向的选择。我们在此描述一种用于修饰CD133干细胞的安全且高效的方法。我们期望这种方法能为优化治疗性干细胞效果以及通过磁共振成像（MRI）监测给药的细胞产品提供一种标准技术。
