Thudium Christian S, Engstrom Amalie, Groen Solveig S, Karsdal Morten A, Bay-Jensen Anne-Christine
Nordic Bioscience;
Nordic Bioscience; Department of Biomedical Sciences, University of Copenhagen.
J Vis Exp. 2019 Nov 3(153). doi: 10.3791/59467.
Ex vivo culture systems cover a broad range of experiments dedicated to studying tissue and cellular function in a native setting. Cartilage is a unique tissue important for proper function of the synovial joint and is constituted by a dense extracellular matrix (ECM), rich in proteoglycan and type II collagen. Chondrocytes are the only cell type present within cartilage and are widespread and relatively low in number. Altered external stimuli and cellular signalling can lead to changes in ECM composition and deterioration, which are important pathological hallmarks in diseases such as osteoarthritis (OA) and rheumatoid arthritis. Ex vivo cartilage models allow 1) profiling of chondrocyte mediated alterations of cartilage tissue turnover, 2) visualizing the cartilage ECM composition, and 3) chondrocyte rearrangement directly in the tissue. Profiling these alterations in response to stimuli or treatments are of high importance in various aspects of cartilage biology, and complement in vitro experiments in isolated chondrocytes, or more complex models in live animals where experimental conditions are more difficult to control. Cartilage explants present a translational and easily accessible method for assessing tissue remodeling in the cartilage ECM in controllable settings. Here, we describe a protocol for isolating and culturing live bovine cartilage explants. The method uses tissue from the bovine knee, which is easily accessible from the local butchery. Both explants and conditioned culture medium can be analyzed to investigate tissue turnover, ECM composition, and chondrocyte function, thus profiling ECM modulation.
体外培养系统涵盖了广泛的实验，致力于在天然环境中研究组织和细胞功能。软骨是一种对滑膜关节正常功能至关重要的独特组织，由富含蛋白聚糖和II型胶原蛋白的致密细胞外基质（ECM）构成。软骨细胞是软骨中唯一存在的细胞类型，分布广泛且数量相对较少。外部刺激和细胞信号的改变可导致ECM组成的变化和退化，这是骨关节炎（OA）和类风湿性关节炎等疾病的重要病理特征。体外软骨模型能够：1）分析软骨细胞介导的软骨组织更新变化；2）观察软骨ECM组成；3）直接在组织中观察软骨细胞重排。分析这些对刺激或处理的反应变化在软骨生物学的各个方面都非常重要，并且补充了在分离的软骨细胞中进行的体外实验，或在实验条件更难控制的活体动物中进行的更复杂模型实验。软骨外植体提供了一种在可控环境中评估软骨ECM中组织重塑的转化且易于获取的方法。在这里，我们描述了一种分离和培养活牛软骨外植体的方案。该方法使用来自牛膝关节的组织，可从当地屠宰场轻松获取。外植体和条件培养基均可进行分析，以研究组织更新、ECM组成和软骨细胞功能，从而分析ECM调节情况。
