Patient-specific induced pluripotent stem (iPS) cells for endothelialisation of membrane surfaces of implantable biohybrid lung devices

用于植入式生物混合肺装置膜表面内皮化的患者特异性诱导多能干 (iPS) 细胞

• 批准号: 347326394
• 负责人: Professor Dr. Ulrich Martin
• 金额: --
• 依托单位: Leibniz Forschungslaboratorien für Biotechnologie und künstliche Organe (LEBAO)
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/347326394?language=en
• 关键词: Patient; specific; induced; pluripotent; stem

One strategy to overcome the limited durability of current lung assist devices due to clotting and thrombus formation is the functionalization of synthetic membrane surfaces. Chemical modification or coating with biological substances such as heparin have been applied to reduce thrombocyte adhesion. Although such strategies can improve the hemocompatibility to a certain extent, long-term functionality of the gas exchange membranes has not been achieved yet. It has been hypothesized that stable endothelialization of blood contacting materials may create a natural non-thrombogenic surface, which could allow for long-term use of membrane oxygenators without thrombus formation.While first studies aiming at the endothelialization of gas exchange membranes show encouraging results, an endothelial cell (EC) source suitable for clinical scale up and application is not yet identified. For seeding of a typical membrane oxygenator with a membrane surface of about 1.3 to 4 sqm, a cell source with high expansion potential will be necessary, that can provide genetically stable ECs with all typical functional properties of cells in the natural EC layer. It is noteworthy that recent developments towards an endothelialized implantable membrane oxygenator did neglect these needs as well as recent reports that demonstrated frequent chromosomal aberrations in cultured primary ECs. Due to the lack of sufficient expandability of adult primary ECs (e.g. from saphena veins), most experimental studies used ECs isolated from human cord blood or umbilical veins.Based on our previous results we hypothesize that patient-specific iPSC-derived ECs represent the most suitable cell source for endothelialization of membrane oxygenators. Here we propose to conduct a systematic study that analyses expandability, genetic integrity as well as functionality of ECs from different cell sources (late outgrowth ECs from cord and adult peripheral blood, ECs from umbilical veins and adult saphena veins, microvascular ECs from adipose tissue as well as endothelial cells differentiated from human iPSCs). In particular, we will investigate senescence-related changes in cell function and secretory characteristics after extended culture expansion that would be required to provide the necessary cell numbers of about 1 to 2 billion cells per device. This project will thus critically contribute to clinical translation of biofunctionalized membrane oxygenators and other EC-based cellular therapies.

克服当前肺辅助装置由于凝血和血栓形成而导致的耐用性有限的一种策略是合成膜表面的功能化。化学修饰或用生物物质（例如肝素）涂覆已用于减少血小板粘附。尽管此类策略可以在一定程度上改善血液相容性，但气体交换膜的长期功能尚未实现。据推测，血液接触材料的稳定内皮化可能会产生天然的非血栓形成表面，从而可以长期使用膜氧合器而不会形成血栓。虽然针对气体交换膜内皮化的初步研究显示出令人鼓舞的结果，但尚未确定适合临床规模化和应用的内皮细胞（EC）来源。对于膜表面约 1.3 至 4 平方米的典型膜氧合器的接种，需要具有高扩增潜力的细胞源，它可以提供具有天然 EC 层细胞的所有典型功能特性的遗传稳定 EC。值得注意的是，最近内皮化植入式膜氧合器的发展确实忽视了这些需求，而且最近的报告表明培养的原代 EC 中经常出现染色体畸变。由于成人原代 EC（例如来自隐静脉）缺乏足够的扩展性，大多数实验研究使用从人脐带血或脐静脉中分离的 EC。根据我们之前的结果，我们假设患者特异性 iPSC 衍生的 EC 代表了最适合膜氧合器内皮化的细胞来源。在这里，我们建议进行一项系统研究，分析来自不同细胞来源的EC（来自脐带和成人外周血的晚期生长EC、来自脐静脉和成人隐静脉的EC、来自脂肪组织的微血管EC以及从人iPSC分化的内皮细胞）的扩展性、遗传完整性和功能。特别是，我们将研究扩展培养扩增后细胞功能和分泌特性的衰老相关变化，这需要为每个设备提供约 1 至 20 亿个细胞的必要细胞数量。因此，该项目将对生物功能化膜氧合器和其他基于 EC 的细胞疗法的临床转化做出重要贡献。