Najimi Mustapha, Smets Françoise, Sokal Etienne
Université Catholique de Louvain, Laboratory of Pediatric Hepatology & Cell Therapy, Brussels, Belgium.
Methods Mol Biol. 2009;481:59-74. doi: 10.1007/978-1-59745-201-4_6.
Apoptosis has been documented as a frequent hurdle phenomenon that occurs in human hepatocytes during isolation, storage, infusion and after engraftment within the recipient liver parenchyma. Apoptosis is an active form of cell death that involves programmed cellular machineries leading to a progressive self-destruction of the cell. In contrary to necrosis, it can affect individual cells within a cell population. It is characterized by chronological alteration of intracellular biochemical signaling pathways followed by cellular morphological changes, DNA fragmentation, perturbation of mitochondrial membrane function and changes in the plasma membrane. These cellular alterations can be analyzed using different methodologies on adherent, suspended and in situ engrafted hepatocytes. This chapter presents a brief overview of these techniques and provides methodology for the evaluation of hepatocyte apoptosis at the structural and biochemical levels.
凋亡已被证明是一种常见的障碍现象,在人肝细胞分离、储存、输注期间以及植入受体肝实质后均会发生。凋亡是一种细胞死亡的主动形式,涉及程序性细胞机制,导致细胞逐渐自我毁灭。与坏死相反,它可影响细胞群体中的单个细胞。其特征是细胞内生化信号通路随时间发生改变,随后出现细胞形态变化、DNA片段化、线粒体膜功能紊乱以及质膜变化。这些细胞改变可以通过不同方法对贴壁、悬浮和原位植入的肝细胞进行分析。本章简要概述了这些技术,并提供了在结构和生化水平评估肝细胞凋亡的方法。