Division of Cardiology, Department of Medicine, Emory University School of Medicine, Atlanta, GA, USA.
Louisiana State University Health Sciences Center, Department of Molecular & Cellular Physiology, Shreveport, LA, USA.
Methods Mol Biol. 2024;2835:155-164. doi: 10.1007/978-1-0716-3995-5_14.
Direct reprogramming provides a novel breakthrough for generating functional endothelial cells (ECs) without the need for intermediate stem or progenitor states, offering a promising resource for cardiovascular research and treatment. ETV2 is a key transcription factor that has been identified as a pioneering factor for specifying endothelial lineage. Achieving precise ETV2 induction is essential for effective endothelial reprogramming, and maintaining the reprogrammed cellular phenotype relies on a specific combination of growth factors and small molecules. Thus, we hereby provide a straightforward and comprehensive protocol for generating two distinct types of reprogrammed ECs (rECs) from human dermal fibroblasts (HDFs). Early rECs demonstrate a robust neovascularization property but lack the mature EC phenotype, while late rECs exhibit phenotypical similarity to human postnatal ECs and have a neovascularization capacity similar to early rECs. Both cell types can be derived from human somatic source cells, making them suitable for personalized disease investigations, drug discovery, and disease therapy.
直接重编程为生成功能性内皮细胞（ECs）提供了一个新的突破，无需中间的干细胞或祖细胞状态，为心血管研究和治疗提供了有前途的资源。ETV2 是一种关键的转录因子，已被确定为指定内皮谱系的先驱因子。实现精确的 ETV2 诱导对于有效的内皮重编程至关重要，而维持重编程的细胞表型依赖于特定的生长因子和小分子组合。因此，我们在此提供了一个简单而全面的方案，用于从人真皮成纤维细胞（HDFs）中生成两种不同类型的重编程 EC（rEC）。早期 rEC 表现出强大的血管生成特性，但缺乏成熟的 EC 表型，而晚期 rEC 表现出与人出生后 EC 相似的表型，并且具有与早期 rEC 相似的血管生成能力。这两种细胞类型都可以从人体体细胞中衍生而来，使其适合于个性化疾病研究、药物发现和疾病治疗。
