Department of Medicine, Surgery and Dentistry "Scuola Medica Salernitana", University of Salerno.
Department of Public Health, University of Naples Federico II.
J Vis Exp. 2024 Mar 22(205). doi: 10.3791/66160.
The myocardium is composed of cardiomyocytes and an even greater number of fibroblasts, the latter being responsible for extracellular matrix production. From the early stages of heart development throughout the lifetime, in both normal and pathological conditions, the composition of the extracellular matrix changes and influences myocardium structure and function. The purpose of the method described here is to obtain the substrate for the culture of cardiac cells in vitro (termed cardiac ECM), mimicking the myocardial extracellular matrix in vivo. To this end, fibroblasts isolated from the adult human heart were cultured to confluence on gelatin-coated dishes to produce the myocardium-specific extracellular matrix. The subsequent removal of cardiac fibroblasts, while preserving the deposited cardiac ECM, produced the substrate for studying the influence of the myocardium-specific extracellular matrix on other cells. Importantly, the composition of the fibroblast-derived coating of the culture dish changes according to the in vivo activity of the fibroblasts isolated from the heart, allowing subsequent studies of cell-matrix interactions in different normal and pathological conditions.
心肌由心肌细胞和数量更多的成纤维细胞组成，后者负责细胞外基质的产生。从心脏发育的早期到整个生命过程中，在正常和病理条件下，细胞外基质的组成都会发生变化，并影响心肌的结构和功能。这里描述的方法旨在获得体外培养心脏细胞的基质（称为心脏 ECM），模拟体内心肌细胞外基质。为此，将从成人心脏分离的成纤维细胞培养至汇合，在明胶包被的培养皿上产生心肌特异性细胞外基质。随后去除心脏成纤维细胞，同时保留沉积的心脏 ECM，得到用于研究心肌特异性细胞外基质对其他细胞影响的基质。重要的是，从心脏分离的成纤维细胞的培养皿的纤维细胞衍生涂层的组成根据其在体内的活性而变化，从而可以在不同的正常和病理条件下进行后续的细胞-基质相互作用研究。
