Severance Integrative Research Institute for Cerebral and Cardiovascular Disease, Yonsei University Health System, Seoul 03722, Republic of Korea.
Department of Bioengineering, College of Engineering, Hanyang University, Seoul 04763, Republic of Korea.
Mol Med Rep. 2018 Dec;18(6):5563-5571. doi: 10.3892/mmr.2018.9561. Epub 2018 Oct 16.
The present study was conducted in order to improve gene expression efficiency of insulin‑like growth factor‑1 (IGF‑1)‑transfected mesenchymal stem cells (MSCs) using a non‑viral carrier and a simplified method of dual gene selection. The therapeutic efficacy of this MSC‑based IGF‑1/enhanced green fluorescent protein (EGFP) dual gene sorting system was evaluated in a rat myocardial infarction (MI) model. IGF‑1 and EGFP genes were expressed in MSCs in vitro. The purity of dual gene‑expressing MSCs was 95.1% by fluorescence‑activated cell sorting. Transfected MSCs injected into rats were identified based on green fluorescence, with an increased signal intensity observed in rats injected with sorted cells, compared with unsorted cells. IGF‑1 expression levels were additionally increased in the sorted group, and decreases in infarct size, fibrotic area and fraction of apoptotic cells were observed. These results demonstrated that IGF‑1 overexpression protects against fibrosis and apoptosis in the myocardium and reduces infarct size following MI. Additionally, the present vector sorting system may potentially be applied to other types of stem cell‑based gene therapy.
本研究旨在通过非病毒载体和简化的双基因选择方法提高转胰岛素样生长因子-1 (IGF-1) 的间充质干细胞 (MSC) 的基因表达效率。采用基于 MSC 的 IGF-1/增强型绿色荧光蛋白 (EGFP) 双基因分选系统在大鼠心肌梗死 (MI) 模型中评估其治疗效果。IGF-1 和 EGFP 基因在 MSC 中体外表达。通过荧光激活细胞分选,双基因表达 MSC 的纯度为 95.1%。注射到大鼠体内的转染 MSC 可通过绿色荧光进行鉴定,与未分选细胞相比,注射分选细胞的大鼠的信号强度增加。分选组 IGF-1 表达水平进一步增加,梗死面积、纤维化面积和凋亡细胞比例降低。这些结果表明 IGF-1 过表达可防止 MI 后心肌纤维化和细胞凋亡,并减小梗死面积。此外,本载体分选系统可能潜在地应用于其他类型的基于干细胞的基因治疗。