Identifying an optimal seeding protocol and endothelial cell substrate for biohybrid lung development
Identifying an optimal seeding protocol and endothelial cell substrate for biohybrid lung development
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DOI：
10.1002/term.2764
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发表时间：
2018-12
影响因子：
3.3
通讯作者：
U. Zwirner;K. Höffler;M. Pflaum;Sotirios Korossis;A. Haverich;B. Wiegmann
中科院分区：
文献类型：
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作者：
U. Zwirner;K. Höffler;M. Pflaum;Sotirios Korossis;A. Haverich;B. Wiegmann
生物杂交肺的发展需要满足几个关键的先决条件，这可以为肺移植提供一种实际的替代方案。一个主要方面是通过内皮细胞种植优化设备人工表面的血液相容性。在这项研究中，四种不同类型的聚合物气体交换中空纤维膜(HFM)使用四种不同的种子方案进行了分析，以确定充分实现长期内皮化的理想组合。用人脐血内皮细胞(HCBECs)内皮化两种不同孔径的聚丙烯和聚4-甲基-1-戊烯-1-戊烯基膜，并对其进行了肝素/白蛋白包覆。通过荧光显微镜、细胞计数和分析活化、切应力和血栓形成状态标志物的相对表达水平，检测了四种不同的旋转种植方案对长期HFM内皮化的定性和定量影响以及炎症刺激对种植的HCBEC的影响。只有使用肝素/白蛋白包被的聚-4-甲基-1-戊烯HFM，在1rpm的转速下旋转24小时，然后静态培养120小时，才能实现优化的内皮细胞接种和长期培养。无论是细胞与人脐静脉内皮细胞接触，还是旋转培养程序，均不影响人脐静脉内皮细胞的生理性、抗血栓形成和抗炎的激活状态。此外，细胞对炎症刺激保持其生理反应。旋转种植策略和无缝的肝素/白蛋白涂层是充分和持久的内皮化的关键要求，因此是未来生物杂交肺开发和体内应用的关键因素。
Several key prerequisites need to be fulfilled for the development of a biohybrid lung, which can offer an actual alternative to lung transplantation. A major aspect is an optimized haemocompatibility of the device's artificial surfaces via endothelial cell seeding. In this study, four different types of polymeric gas exchange hollow fibre membranes (HFMs) were analysed utilizing four different seeding protocols in order to identify the ideal combination for sufficient long‐term endothelialization. Human cord blood‐derived endothelial cells (HCBECs) were used for the endothelialization of polypropylene HFMs with two different pore sizes and poly‐4‐methyl‐1‐pentene HFMs, both with and without heparin/albumin coating. The qualitative and quantitative impact of four different rotational seeding protocols regarding long‐term HFM endothelialization and the impact of inflammatory stimulation on the seeded HCBECs were examined by fluorescence microscopy, cell counting, and analysis of relative expression levels of activation, shear stress, and thrombogenic state markers. Optimized endothelial cell seeding and long‐term cultivation were only achieved using heparin/albumin‐coated poly‐4‐methyl‐1‐pentene HFMs, applying 24 hr of rotational speed at 1 rpm followed by 120 hr of static culture. Neither cell‐to‐HFM contact nor the rotational cultivation procedure showed an impact on the physiological anti‐thrombogenic and anti‐inflammatory HCBEC activation status. Additionally, the cells maintained their physiological responsiveness towards inflammatory stimulation. Rotational seeding strategies and a seamless heparin/albumin coating of the HFMs are crucial requirements for a sufficient and long‐lasting endothelialization and thus a key element in the future development and in vivo application of the biohybrid lung.