Vinken Mathieu, Maes Michaël, Crespo Yanguas Sara, Willebrords Joost, Vanhaecke Tamara, Rogiers Vera
Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090, Brussels, Belgium,
Methods Mol Biol. 2015;1250:95-103. doi: 10.1007/978-1-4939-2074-7_6.
Fas-mediated apoptosis underlies a plethora of liver pathologies and toxicities. As a consequence, this process is a major research topic in the field of experimental and clinical hepatology. The present chapter describes the setup of an in vitro model of hepatocellular apoptotic cell death. In essence, this system consists of freshly isolated hepatocytes cultured in a monolayer configuration, which are exposed to a combination of Fas ligand and cycloheximide. This in vitro model has been characterized by using a set of well-acknowledged cell death markers. This experimental system allows the study of the entire course of Fas-mediated hepatocellular cell death, going from early apoptosis to secondary necrosis, and hence can serve a broad range of in vitro pharmaco-toxicological purposes.
Fas介导的细胞凋亡是众多肝脏病理和毒性的基础。因此，这一过程是实验性和临床肝脏病学领域的一个主要研究课题。本章描述了肝细胞凋亡性细胞死亡体外模型的建立。实质上，该系统由以单层形式培养的新鲜分离的肝细胞组成，这些肝细胞暴露于Fas配体和放线菌酮的组合中。该体外模型已通过一组公认的细胞死亡标志物进行了表征。这个实验系统可以研究Fas介导的肝细胞死亡的整个过程，从早期凋亡到继发性坏死，因此可用于广泛的体外药物毒理学研究目的。
