Institute of Pharmacology and Toxicology, University Medical Center Goettingen; DZHK (German Center for Cardiovascular Research) partner site, Goettingen;
Institute of Pharmacology and Toxicology, University Medical Center Goettingen; DZHK (German Center for Cardiovascular Research) partner site, Goettingen.
J Vis Exp. 2021 Aug 20(174). doi: 10.3791/62700.
Fibroblasts are phenotypically highly dynamic cells, which quickly transdifferentiate into myofibroblasts in response to biochemical and biomechanical stimuli. The current understanding of fibrotic processes, including cardiac fibrosis, remains poor, which hampers the development of new anti-fibrotic therapies. Controllable and reliable human model systems are crucial for a better understanding of fibrosis pathology. This is a highly reproducible and scalable protocol to generate engineered connective tissues (ECT) in a 48-well casting plate to facilitate studies of fibroblasts and the pathophysiology of fibrotic tissue in a 3-dimensional (3D) environment. ECT are generated around the poles with tunable stiffness, allowing for studies under a defined biomechanical load. Under the defined loading conditions, phenotypic adaptations controlled by cell-matrix interactions can be studied. Parallel testing is feasible in the 48-well format with the opportunity for the time-course analysis of multiple parameters, such as tissue compaction and contraction against the load. From these parameters, biomechanical properties such as tissue stiffness and elasticity can be studied.
成纤维细胞是表型高度动态的细胞，它们会迅速向肌成纤维细胞转分化，以响应生化和生物力学刺激。目前对纤维化过程（包括心脏纤维化）的理解仍然很差，这阻碍了新的抗纤维化治疗方法的发展。可控且可靠的人类模型系统对于更好地理解纤维化病理学至关重要。这是一种在 48 孔铸造板中生成工程化结缔组织 (ECT) 的高度可重复和可扩展的方案，可促进在 3 维 (3D) 环境中研究成纤维细胞和纤维化组织的病理生理学。ECT 围绕着两极生成，具有可调的刚度，允许在定义的生物力学负载下进行研究。在定义的加载条件下，可以研究由细胞-基质相互作用控制的表型适应。在 48 孔格式中可以进行并行测试，有机会对多个参数（如组织压实和对负载的收缩）进行时间过程分析。从这些参数中，可以研究组织刚度和弹性等生物力学特性。
