Bors Luca Anna, Maczelka Hédi, Merkely Béla, Apáti Ágota, Kriston-Vizi Janos, Földes Gábor
Heart and Vascular Center, Semmelweis University, Budapest, Hungary.
Institute of Molecular Life Sciences, HUN-REN Research Centre for Natural Sciences, Budapest, Hungary.
Methods Mol Biol. 2025;2924:113-130. doi: 10.1007/978-1-0716-4530-7_9.
Endothelial dysfunction plays a critical role in the pathophysiology of numerous cardiovascular diseases, the leading cause of mortality globally. Accurate endothelial models are essential for disease modeling, understanding mechanisms of disease development, and drug screening. Animal models, though informative, often fail to replicate human disease conditions, as well as ethical considerations advocate for reducing animal experimentation in drug research. Human primary endothelial cells offer mature endothelial characteristics but are limited by availability, complicating reproducibility. Human pluripotent stem cell-derived endothelial cells (hPSC-ECs) emerge as a promising alternative, providing an unlimited cell source. This study presents multiple useful methods to evaluate the phenotypic and functional properties of hiPSC-ECs. Immunohistochemistry and fluorescence-activated cell sorting (FACS) validated endothelial characteristics, morphology, texture, and marker presence, ensuring differentiation efficacy and cell culture viability. Functional assays, including wound healing and 3D spheroid-based angiogenesis, demonstrated that hiPSC-ECs mimic native endothelial cells. High-content screening approaches analyzed the endothelial phenotype, highlighting the potential of hiPSC-ECs in cardiovascular disease research and drug development.
内皮功能障碍在众多心血管疾病的病理生理学中起着关键作用，而心血管疾病是全球死亡的主要原因。准确的内皮模型对于疾病建模、理解疾病发展机制以及药物筛选至关重要。动物模型虽然具有参考价值，但往往无法复制人类疾病状况，而且出于伦理考虑，在药物研究中提倡减少动物实验。人原代内皮细胞具有成熟的内皮特征，但受到可用性的限制，使得可重复性变得复杂。人多能干细胞衍生的内皮细胞（hPSC-ECs）作为一种有前景的替代方案出现，提供了无限的细胞来源。本研究提出了多种有用的方法来评估hiPSC-ECs的表型和功能特性。免疫组织化学和荧光激活细胞分选（FACS）验证了内皮特征、形态、质地和标志物的存在，确保了分化效率和细胞培养活力。包括伤口愈合和基于3D球体的血管生成在内的功能测定表明，hiPSC-ECs模拟天然内皮细胞。高内涵筛选方法分析了内皮表型，突出了hiPSC-ECs在心血管疾病研究和药物开发中的潜力。
