Acellular Normal and Fibrotic Human Lung Matrices as a Culture System for In Vitro Investigation
Acellular Normal and Fibrotic Human Lung Matrices as a Culture System for In Vitro Investigation
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DOI：
10.1164/rccm.201204-0754oc
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发表时间：
2012-11-01
影响因子：
24.7
通讯作者：
White, Eric S.
中科院分区：
文献类型：
--
作者：
Booth, Adam J.;Hadley, Ryan;White, Eric S.
基本原理：细胞外基质（ECM）是一种动态的组织，有助于器官的完整性和功能，其对细胞表型的调节是细胞生物学的一个主要方面。然而，标准的体外培养方法的生理相关性尚不清楚，因为它们不能模拟活体器官的组成、结构或可扩张性。在肺，成纤维细胞存在于ECM丰富的间隙空间，是肺fibrogenesis.Objectives的关键效应子：为了更好地解决ECM如何影响成纤维细胞表型的疾病特异性的方式，我们开发了一种培养系统，使用脱细胞人类正常和fibrotic lungs.Methods：脱细胞是使用洗涤剂，盐，和DNase治疗实现。所得的矩阵可以被分割为均匀的切片内的细胞culture.Measurements和主要结果：我们报告说，脱细胞过程中有效地去除细胞和核材料，同时保留本机的尺寸和刚度的肺组织。我们证明，肺成纤维细胞restressed到脱细胞肺基质可以随后使用常规协议进行分析，以这种方式，我们表明，纤维化基质明显促进转化生长因子-β-非依赖性肌成纤维细胞分化与正常基质相比。此外，全面分析的无细胞基质ECM的细节显着组成正常和纤维化肺之间的差异，铺平了道路，为进一步研究的新hypothesis.Conclusions：这种方法有望使重要的ECM为基础的假说在人体组织中的调查，并允许未来的科学探索在器官和疾病的具体方式。
Rationale: Extracellular matrix (ECM) is a dynamic tissue that contributes to organ integrity and function, and its regulation of cell phenotype is a major aspect of cell biology. However, standard in vitro culture approaches are of unclear physiologic relevance because they do not mimic the compositional, architectural, or distensible nature of a living organ. In the lung, fibroblasts exist in ECM-rich interstitial spaces and are key effectors of lung fibrogenesis.Objectives: To better address how ECM influences fibroblast phenotype in a disease-specific manner, we developed a culture system using acellular human normal and fibrotic lungs.Methods: Decellularization was achieved using treatment with detergents, salts, and DNase. The resultant matrices can be sectioned as uniform slices within which cells were cultured.Measurements and Main Results: We report that the decellularization process effectively removes cellular and nuclear material while retaining native dimensionality and stiffness of lung tissue. We demonstrate that lung fibroblasts reseeded into acellular lung matrices can be subsequently assayed using conventional protocols; in this manner we show that fibrotic matrices clearly promote transforming growth factor-beta-independent myofibroblast differentiation compared with normal matrices. Furthermore, comprehensive analysis of acellular matrix ECM details significant compositional differences between normal and fibrotic lungs, paving the way for further study of novel hypotheses.Conclusions: This methodology is expected to allow investigation of important ECM-based hypotheses in human tissues and permits future scientific exploration in an organ- and disease-specific manner.