Bölükbas Deniz A, De Santis Martina M, Alsafadi Hani N, Doryab Ali, Wagner Darcy E
Department of Experimental Medical Sciences, Faculty of Medicine, Lund University, Lund, Sweden.
Wallenberg Centre for Molecular Medicine, Faculty of Medicine, Lund University, Lund, Sweden.
Methods Mol Biol. 2019;1940:275-295. doi: 10.1007/978-1-4939-9086-3_20.
Lung transplantation is the only option for patients with end-stage lung disease, but there is a shortage of available lung donors. Furthermore, efficiency of lung transplantation has been limited due to primary graft dysfunction. Recent mouse models mimicking lung disease in humans have allowed for deepening our understanding of disease pathomechanisms. Moreover, new techniques such as decellularization and recellularization have opened up new possibilities to contribute to our understanding of the regenerative mechanisms involved in the lung. Stripping the lung of its native cells allows for unprecedented analyses of extracellular matrix and sets a physiologic platform to study the regenerative potential of seeded cells. A comprehensive understanding of the molecular pathways involved for lung development and regeneration in mouse models can be translated to regeneration strategies in higher organisms, including humans. Here we describe and discuss several techniques used for murine lung de- and recellularization, methods for evaluation of efficacy including histology, protein/RNA isolation at the whole lung, as well as lung slices level.
肺移植是终末期肺病患者的唯一选择,但可用的肺供体短缺。此外,由于原发性移植功能障碍,肺移植的效率一直受到限制。最近模拟人类肺病的小鼠模型加深了我们对疾病发病机制的理解。此外,去细胞化和再细胞化等新技术为我们理解肺的再生机制开辟了新的可能性。去除肺的天然细胞可以对细胞外基质进行前所未有的分析,并为研究接种细胞的再生潜力提供一个生理平台。对小鼠模型中肺发育和再生所涉及的分子途径的全面理解可以转化为高等生物(包括人类)的再生策略。在这里,我们描述并讨论了几种用于小鼠肺去细胞化和再细胞化的技术,包括组织学、全肺及肺切片水平的蛋白质/RNA分离等评估功效的方法。