Cell and Gene Therapy, Departments of Biomedicine and Surgery, University and University Hospital Basel, Basel, Switzerland.
Cardiac Surgery and Engineering, Departments of Biomedicine and Surgery, University and University Hospital Basel, Basel, Switzerland.
J Cell Mol Med. 2018 May;22(5):2580-2591. doi: 10.1111/jcmm.13511. Epub 2018 Feb 25.
Vascular Endothelial Growth Factor (VEGF) can induce normal or aberrant angiogenesis depending on the amount secreted in the microenvironment around each cell. Towards a possible clinical translation, we developed a Fluorescence Activated Cell Sorting (FACS)-based technique to rapidly purify transduced progenitors that homogeneously express a desired specific VEGF level from heterogeneous primary populations. Here, we sought to induce safe and functional angiogenesis in ischaemic myocardium by cell-based expression of controlled VEGF levels. Human adipose stromal cells (ASC) were transduced with retroviral vectors and FACS purified to generate two populations producing similar total VEGF doses, but with different distributions: one with cells homogeneously producing a specific VEGF level (SPEC), and one with cells heterogeneously producing widespread VEGF levels (ALL), but with an average similar to that of the SPEC population. A total of 70 nude rats underwent myocardial infarction by coronary artery ligation and 2 weeks later VEGF-expressing or control cells, or saline were injected at the infarction border. Four weeks later, ventricular ejection fraction was significantly worsened with all treatments except for SPEC cells. Further, only SPEC cells significantly increased the density of homogeneously normal and mature microvascular networks. This was accompanied by a positive remodelling effect, with significantly reduced fibrosis in the infarcted area. We conclude that controlled homogeneous VEGF delivery by FACS-purified transduced ASC is a promising strategy to achieve safe and functional angiogenesis in myocardial ischaemia.
血管内皮生长因子（VEGF）可根据其在细胞周围微环境中分泌的量，诱导正常或异常的血管生成。为了可能的临床转化，我们开发了一种基于荧光激活细胞分选（FACS）的技术，以快速从异质原代群体中纯化均匀表达所需特定 VEGF 水平的转导祖细胞。在这里，我们试图通过基于细胞的表达来控制 VEGF 水平，从而在缺血性心肌中诱导安全和功能性血管生成。用逆转录病毒载体转导人脂肪基质细胞（ASC），并通过 FACS 进行纯化，以产生两种产生相似总 VEGF 剂量但具有不同分布的群体：一种是均匀产生特定 VEGF 水平的细胞（SPEC），另一种是不均匀产生广泛的 VEGF 水平的细胞（ALL），但平均水平与 SPEC 群体相似。共有 70 只裸鼠通过冠状动脉结扎术发生心肌梗死，2 周后在梗死边界注射表达 VEGF 的或对照细胞或生理盐水。4 周后，除 SPEC 细胞外，所有治疗均显著降低心室射血分数。此外，只有 SPEC 细胞显著增加了均匀正常和成熟微血管网络的密度。这伴随着积极的重塑效应，梗死区的纤维化明显减少。我们得出结论，通过 FACS 纯化的转导 ASC 进行的受控均匀 VEGF 输送是在心肌缺血中实现安全和功能性血管生成的有前途的策略。
