Künzel Stephan R, Schaeffer Charlotte, Sekeres Karolina, Mehnert Carola S, Schacht Wall Stephanie M, Newe Manja, Kämmerer Susanne, El-Armouche Ali
Institute of Pharmacology and Toxicology, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden;
Institute of Pharmacology and Toxicology, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden.
J Vis Exp. 2019 Jul 29(149). doi: 10.3791/59858.
Primary adult fibroblasts have become an important tool to study fibrosis, fibroblast interactions and inflammation in all body tissues. Since primary fibroblasts cannot divide indefinitely due to myofibroblast differentiation or senescence induction, new cultures must be established regularly. However, there are several obstacles to overcome during the processes of developing a reliable isolation protocol and primary fibroblast isolation itself: the method's degree of difficulty (especially for beginners), the risk of bacterial contamination, the required time until primary fibroblasts can be used for experiments, and subsequent cell quality and viability. In this study, a fast, reliable and easy-to-learn protocol to isolate and culture primary adult fibroblasts from mouse heart, lung, liver and kidney combining enzymatic digestion and ultrasonic agitation is provided.
原代成人成纤维细胞已成为研究所有身体组织中的纤维化、成纤维细胞相互作用和炎症的重要工具。由于原代成纤维细胞会因肌成纤维细胞分化或衰老诱导而无法无限分裂，因此必须定期建立新的培养物。然而，在制定可靠的分离方案和原代成纤维细胞分离过程本身中，有几个障碍需要克服：该方法的难度（尤其是对于初学者）、细菌污染的风险、直到原代成纤维细胞可用于实验所需的时间，以及随后的细胞质量和活力。在本研究中，提供了一种快速、可靠且易于学习的方案，该方案结合酶消化和超声搅拌从小鼠心脏、肺、肝脏和肾脏中分离和培养原代成人成纤维细胞。
