Department of Medical Physiology, Texas A&M University College of Medicine, Brain, TX, USA.
Methods Mol Biol. 2021;2319:143-152. doi: 10.1007/978-1-0716-1480-8_16.
Heart disease is one of the leading causes of death in the United States. Isolation and culture adult cardiomyocytes are important for studying cardiomyocyte contractility, heart hypertrophy, and cardiac failure. In contrast to neonatal cardiomyocyte isolation, adult mice cardiomyocytes isolation is challenging due to firm connections among cardiomyocytes through intercalated discs. The availability of newly generated genetically modified mouse lines requires to establish protocols to isolation and culture adult mouse cardiomyocyte for in vitro studies. In this manuscript, we described a straightforward method of isolating adult mouse cardiomyocytes using Langendorff perfusion apparatus. Briefly, the hearts were harvested from adult mice and the heart was mounted to Lagendorff apparatus. After perfusion with calcium depletion and collagenase digestion, the left ventricles were minced and filtered. Lastly, the separated cardiomyocytes were treated with CaCl. The isolated cardiac myocytes can be utilized in a broad range of experiments including screening for drugs.
心脏病是美国的主要死因之一。分离和培养成人心肌细胞对于研究心肌细胞收缩性、心脏肥大和心力衰竭非常重要。与新生大鼠心肌细胞分离不同，由于心肌细胞通过闰盘紧密连接，成年小鼠心肌细胞的分离具有挑战性。新产生的基因修饰小鼠品系的可用性需要建立分离和培养成年小鼠心肌细胞的方案，用于体外研究。在本手稿中，我们描述了一种使用 Langendorff 灌注装置分离成年小鼠心肌细胞的简单方法。简而言之，从成年小鼠中取出心脏，并将其安装到 Langendorff 装置上。在用钙耗竭和胶原酶消化后，将左心室切碎并过滤。最后，用 CaCl 处理分离的心肌细胞。分离的心肌细胞可用于广泛的实验，包括药物筛选。
