A Pulmonary Vascular Model From Endothelialized Whole Organ Scaffolds.
A Pulmonary Vascular Model From Endothelialized Whole Organ Scaffolds.
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DOI：
10.3389/fbioe.2021.760309
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发表时间：
2021
影响因子：
5.7
通讯作者：
Niklason LE
中科院分区：
文献类型：
--
作者：
Yuan Y;Leiby KL;Greaney AM;Raredon MSB;Qian H;Schupp JC;Engler AJ;Baevova P;Adams TS;Kural MH;Wang J;Obata T;Yoder MC;Kaminski N;Niklason LE
研究肺血管疾病的体外系统的发展对于理解人类的病理学至关重要。传统的培养系统不能完全概括当地的微环境条件，而且其代表人类病理生理学研究疾病和药物机制的能力通常是有限的。整个器官脱细胞提供了一种方法来开发一种结构，该结构概括了完整的血管结构的结构、机械和生物学特征。在这里，我们开发了一种培养方案来提高全肺支架中内皮细胞的覆盖率，并使用单细胞RNA测序分析来探索在仿生生物反应器系统中去细胞全肺支架对内皮细胞表型和功能的影响。有趣的是，我们发现，原始肺微血管的表型和功能信号至少部分回复到天然肺内皮细胞。此外，在去细胞肺系统中培养的人诱导多能干细胞来源的内皮细胞开始获得各种天然的人内皮细胞表型。血管屏障功能部分恢复，而内皮细胞重植肺的小毛细血管仍然通畅。为了评价工程化内皮细胞对外源性刺激的通透性调节能力，将脂多糖(LPS)引入再生肺，模拟急性肺损伤。内毒素治疗后，促炎信号明显增强，血管屏障受损。综上所述，这些结果展示了一个新的平台，在整个器官水平上概括了一些肺微血管功能和表型。这一进展可能有助于为使用整体器官工程学方法来模拟血管疾病铺平道路。
The development of an in vitro system for the study of lung vascular disease is critical to understanding human pathologies. Conventional culture systems fail to fully recapitulate native microenvironmental conditions and are typically limited in their ability to represent human pathophysiology for the study of disease and drug mechanisms. Whole organ decellularization provides a means to developing a construct that recapitulates structural, mechanical, and biological features of a complete vascular structure. Here, we developed a culture protocol to improve endothelial cell coverage in whole lung scaffolds and used single-cell RNA-sequencing analysis to explore the impact of decellularized whole lung scaffolds on endothelial phenotypes and functions in a biomimetic bioreactor system. Intriguingly, we found that the phenotype and functional signals of primary pulmonary microvascular revert back—at least partially—toward native lung endothelium. Additionally, human induced pluripotent stem cell-derived endothelium cultured in decellularized lung systems start to gain various native human endothelial phenotypes. Vascular barrier function was partially restored, while small capillaries remained patent in endothelial cell-repopulated lungs. To evaluate the ability of the engineered endothelium to modulate permeability in response to exogenous stimuli, lipopolysaccharide (LPS) was introduced into repopulated lungs to simulate acute lung injury. After LPS treatment, proinflammatory signals were significantly increased and the vascular barrier was impaired. Taken together, these results demonstrate a novel platform that recapitulates some pulmonary microvascular functions and phenotypes at a whole organ level. This development may help pave the way for using the whole organ engineering approach to model vascular diseases.