In vitro organization of human induced pluripotent stem cell-derived cardiovascular cells into vascularized cardiac tissue

人诱导多能干细胞来源的心血管细胞体外组织成血管化心脏组织

• 批准号: 242448821
• 负责人: Professorin Dr. Ina Gruh
• 金额: --
• 依托单位: Klinik für Herz-, Thorax-, Transplantations- und Gefäßchirurgie (HTTG)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/242448821?language=en
• 关键词: vitro; organization; human; induced; pluripotent

Cardiac tissue engineering strategies may provide new therapeutic options for the treatment of cardiovascular disease, with patient-specific stem-cell derived cardiovascular cells as an ideal cell source. For the in vitro generation of functional heart tissue, sufficient vascularization by endothelial cells and smooth muscle cells is a critical aspect in addition to the contractile force of cardiomyocytes and mechanical stabilization by fibroblasts. Recent findings offer the unique opportunity to generate all cell types of the heart from pluripotent stem cells. The overall objective of the proposed research is the investigation of mechanisms of stem cell-derived human vascularized bioartificial cardiac tissue formation and self-organization in vitro and its regenerative potential in vivo. We hypothesize that 1) cardiovascular cell types can be derived from a single cell source of human induced pluripotent stem cells (iPSCs); and 2) these cells are able to self-organize in vitro to form fully autologous vascularized myocardial tissue for therapeutic application; and 3) pericytes might play an important role to support the formation of functional vascular structures. Consequently, the specific objectives include:- Optimization of differentiation and/or selection strategies to generate from a single stem cell source (human pluripotent stem cells) five different cell types present in the native heart tissue (cardiomyocytes, endothelial cells, smooth muscle cells, pericytes, and fibroblasts). So far, no single differentiation protocol is available for the generation of all cardiovascular cell types of the heart in a simultaneous one pot reaction, yielding defined and appropriate proportions of the desired cell types. Therefore, the use of individual protocols for the directed differentiation of the respective cell types seems more promising. - Implementation of all cardiovascular cell types for the in vitro generation of functional myocardial tissue. Mechanisms of tissue formation, optimal cell type stoichiometry and spatial organization as well as the need for further in situ maturation will be investigated.- Investigation of the role of pericytes for vascularization in the in vitro model of bioartificial cardiac tissue formation, including analysis of pericyte-derived pro-angiogenic factor expression and activation of cell signaling pathways in other cardiovascular cell types. - Investigation of the regenerative potential of vascularized bioartificial cardiac tissue in vivo in a small animal model of myocardial infarction in guinea-pigs with a direct comparison to non-vascularized tissue.Together with progress concerning patient-specific iPSC generation and their clinical applicability, knowledge gained in this project would facilitate the generation of autologous bioartificial vascularized cardiac tissue from human iPSCs for future tissue replacement therapy of damaged and diseased myocardium.

心脏组织工程策略可能为心血管疾病的治疗提供新的治疗选择，患者特异性干细胞衍生的心血管细胞是理想的细胞来源。对于功能性心脏组织的体外生成，除了心肌细胞的收缩力和成纤维细胞的机械稳定性之外，内皮细胞和平滑肌细胞的充分血管化是关键方面。最近的研究结果提供了从多能干细胞产生心脏所有细胞类型的独特机会。该研究的总体目标是研究干细胞衍生的人血管化生物人工心脏组织的体外形成和自组织机制及其体内再生潜力。我们假设：1）心血管细胞类型可以来源于人诱导多能干细胞（iPSC）的单细胞来源; 2）这些细胞能够在体外自组织形成完全自体血管化的心肌组织用于治疗应用; 3）周细胞可能在支持功能性血管结构的形成方面发挥重要作用。- 优化分化和/或选择策略以从单一干细胞来源（人多能干细胞）产生存在于天然心脏组织中的五种不同细胞类型（心肌细胞、内皮细胞、平滑肌细胞、周细胞和成纤维细胞）。到目前为止，没有单一的分化方案可用于在同时的一锅反应中产生心脏的所有心血管细胞类型，从而产生限定的和适当比例的所需细胞类型。因此，使用个体方案来定向分化相应的细胞类型似乎更有希望。- 实现所有心血管细胞类型用于功能性心肌组织的体外生成。将研究组织形成，最佳细胞类型化学计量和空间组织的机制以及进一步原位成熟的需要。在生物人工心脏组织形成的体外模型中研究周细胞在血管形成中的作用，包括分析周细胞衍生的促血管生成因子表达和其他心血管细胞类型中细胞信号通路的激活。 - 在豚鼠心肌梗死的小动物模型中研究血管化生物人工心脏组织的体内再生潜力，并与非血管化组织进行直接比较。该项目中获得的知识将有助于从人类iPSC中产生自体生物人工血管化心脏组织，用于未来的组织替代治疗受损和患病的心肌。