Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, REBIRTH-Cluster of Excellence, Hannover Medical School, Carl-Neuberg-Str.1, 30625 Hannover, Germany.
Eur Heart J. 2011 Nov;32(21):2634-41. doi: 10.1093/eurheartj/ehr166. Epub 2011 May 19.
Induced pluripotent stem cell (iPSC)-derived cardiovascular progenitor cells represent a suitable autologous cell source for myocardial regeneration as they have the capability to form myocardial cells and to contribute to revascularization. As a first proof of concept we evaluated the potential of a murine iPSC-derived cardiovascular progenitor population, which expresses the surface marker foetal liver kinase-1 (Flk-1), to restore myocardial tissue and improve cardiac function after acute myocardial infarction (MI) in mice.
iPSC-derived Flk-1(pos) vs. Flk-1(neg) cells were selected by fluorescence activated cell sorting (FACS) and injected into the ischaemic myocardium of left anterior descending coronary artery (LAD)-ligated mice. Addressing safety aspects we used an octamer binding factor 4 (Oct4)-enhanced green fluorescent protein (eGFP) expressing iPSC clone from the transgenic Oct4-eGFP reporter mouse strain OG2 to enable FACS-based depletion of undifferentiated cells prior to transplantation. Infarcted animals were treated with placebo (phosphate-buffered saline, n = 13), Flk-1(neg) cells (n = 14), or Flk-1(pos) cells (n = 11; 5 × 10(5) cells each). Heart function was evaluated by magnetic resonance imaging and conductance catheter analysis 2 weeks postoperatively. Cardiovascular in vitro and in vivo differentiations were investigated by immunofluorescence staining. Treatment with Flk-1(pos) and Flk-1(neg) cells resulted in a favourable myocardial remodelling and improved left ventricular function. Engraftment and functional benefits were superior after transplantation of Flk-1(pos) compared with Flk-1(neg) cells. Furthermore, Flk-1(pos) grafts contained considerably more vascular structures in relation to Flk-1(neg) grafts.
iPSC-derived Flk-1(pos) progenitor cells differentiate into cardiovascular lineages in vitro and in vivo and improve cardiac function after acute MI. This proof of concept study paves the way for an autologous iPSC-based therapy of MI.
诱导多能干细胞（iPSC）衍生的心血管祖细胞是一种适合心肌再生的自体细胞来源，因为它们具有形成心肌细胞和促进血管生成的能力。作为第一个概念验证，我们评估了表达胎肝激酶 1（Flk-1）表面标志物的鼠 iPSC 衍生心血管祖细胞群体在急性心肌梗死（MI）后恢复心肌组织和改善心脏功能的潜力在结扎左前降支冠状动脉（LAD）的小鼠的心肌中。为了解决安全性问题，我们使用了来自转基因 Oct4-eGFP 报告小鼠品系 OG2 的八聚体结合因子 4（Oct4）增强型绿色荧光蛋白（eGFP）表达 iPSC 克隆，以便在移植前通过荧光激活细胞分选（FACS）基于细胞的耗竭未分化细胞。将梗死动物用安慰剂（磷酸盐缓冲盐水，n = 13）、Flk-1（neg）细胞（n = 14）或 Flk-1（pos）细胞（n = 11; 每次 5×10（5）个细胞）处理。术后 2 周通过磁共振成像和电导导管分析评估心脏功能。通过免疫荧光染色研究心血管体外和体内分化。Flk-1（pos）和 Flk-1（neg）细胞的治疗导致心肌重塑和左心室功能改善。与 Flk-1（neg）细胞相比，Flk-1（pos）细胞移植后的嵌合和功能益处更高。此外，Flk-1（pos）移植物中含有与 Flk-1（neg）移植物相比数量更多的血管结构。
iPSC 衍生的 Flk-1（pos）祖细胞在体外和体内分化为心血管谱系，并改善急性 MI 后的心脏功能。这项概念验证研究为 MI 的自体 iPSC 治疗铺平了道路。
