Development and characterization of a 3D multicell microtissue culture model of airway smooth muscle
Development and characterization of a 3D multicell microtissue culture model of airway smooth muscle
复制标题
DOI：
10.1152/ajplung.00168.2012
复制
发表时间：
2013-01-01
影响因子：
4.9
通讯作者：
Maksym, Geoffrey N.
中科院分区：
文献类型：
--
作者：
West, Adrian R.;Zaman, Nishat;Maksym, Geoffrey N.
West AR、Zaman N、Cole DJ、Walker MJ、Legant WR、Boudou T、Chen CS、Favreau JT、Gaudette GR、Cowley EA、Maksym GN。气道平滑肌 3D 多细胞微组织培养模型的开发和表征。 Am J Physiol Lung Cell Mol Physiol 304：L4-L16，2013 年。首次发表于 2012 年 11 月 2 日； doi:10.1152/ajplung.00168.2012.-气道平滑肌 (ASM) 细胞和分子生物学通常使用生长在平面 2D 基质上的单细胞培养物进行研究。然而，体内细胞作为复杂 3D 结构的一部分存在，并且在其他细胞类型中已充分证实，改变基质几何形状会对表型和功能产生有效影响。这些因素可能与哮喘（一种以气道壁结构重塑为特征的疾病）尤其相关，并强调需要更多生理相关的 ASM 功能模型。我们利用称为微制造组织测量仪的组织工程平台开发了 ASM 3D 培养模型，该模型具有类似于 0.4 毫米长、类似于 350 个细胞“微组织”的阵列，能够同时测量收缩力和细胞级显微镜。仅 ASM 的微组织产生基线张力，表现出强大的细胞组织，并形成肌动蛋白应力纤维，但在 3 天内失去结构完整性并与悬臂分离。 3T3-成纤维细胞的添加显着提高了存活时间，且不影响张力发展或形态。 ASM-3T3 微组织的收缩与离体 ASM 类似，对一系列收缩剂和松弛剂表现出可重复的反应。与 2D 培养物相比，微组织对乙酰胆碱和 KCl 表现出相同的反应，但对组胺、毛喉素或细胞松弛素 D 则不然，这表明收缩性受底物几何形状调节。微组织代表了一种研究 ASM 的新颖模型，它结合了生理 3D 结构、真实的机械环境、多种细胞类型的共培养以及与现有模型相当的收缩特性。这种新模型可以快速筛查生化和机械因素，以深入了解哮喘中的 ASM 功能障碍。
West AR, Zaman N, Cole DJ, Walker MJ, Legant WR, Boudou T, Chen CS, Favreau JT, Gaudette GR, Cowley EA, Maksym GN. Development and characterization of a 3D multicell microtissue culture model of airway smooth muscle. Am J Physiol Lung Cell Mol Physiol 304: L4-L16, 2013. First published November 2, 2012; doi:10.1152/ajplung.00168.2012.-Airway smooth muscle (ASM) cellular and molecular biology is typically studied with single-cell cultures grown on flat 2D substrates. However, cells in vivo exist as part of complex 3D structures, and it is well established in other cell types that altering substrate geometry exerts potent effects on phenotype and function. These factors may be especially relevant to asthma, a disease characterized by structural remodeling of the airway wall, and highlights a need for more physiologically relevant models of ASM function. We utilized a tissue engineering platform known as microfabricated tissue gauges to develop a 3D culture model of ASM featuring arrays of similar to 0.4 mm long, similar to 350 cell "microtissues" capable of simultaneous contractile force measurement and cell-level microscopy. ASM-only microtissues generated baseline tension, exhibited strong cellular organization, and developed actin stress fibers, but lost structural integrity and dissociated from the cantilevers within 3 days. Addition of 3T3-fibroblasts dramatically improved survival times without affecting tension development or morphology. ASM-3T3 microtissues contracted similarly to ex vivo ASM, exhibiting reproducible responses to a range of contractile and relaxant agents. Compared with 2D cultures, microtissues demonstrated identical responses to acetylcholine and KCl, but not histamine, forskolin, or cytochalasin D, suggesting that contractility is regulated by substrate geometry. Microtissues represent a novel model for studying ASM, incorporating a physiological 3D structure, realistic mechanical environment, coculture of multiple cells types, and comparable contractile properties to existing models. This new model allows for rapid screening of biochemical and mechanical factors to provide insight into ASM dysfunction in asthma.