Universidad Pablo de Olavide, Sevilla, Spain.
Redox Biol. 2015 Aug;5:416. doi: 10.1016/j.redox.2015.09.020. Epub 2015 Dec 30.
Apoptosis is characterized by degradation of cell components but plasma membrane remains intact. Apoptotic microtubule network (AMN) is organized during apoptosis forming a cortical structure beneath plasma membrane that maintains plasma membrane integrity. Apoptotic cells are also characterized by high reactive oxygen species (ROS) production that can be potentially harmful for the cell. The aim of this study was to develop a method that allows stabilizing apoptotic cells for diagnostic and therapeutic applications. We were able by using a cocktail composed of taxol (a microtubule stabilizer), Zn (a caspase inhibitor) and coenzyme Q (a lipid antioxidant) to stabilize H460 apoptotic cells in cell cultures for at least 72hours preventing secondary necrosis. Stabilized apoptotic cells maintain many apoptotic cells characteristics such as the presence of apoptotic microtubules, plasma membrane integrity, low intracellular calcium levels, plasma membrane potential, PS externalization and ability of being phagocytosed. Stabilized apoptotic cells can be considered as dying cells in which the cellular cortex and plasma membrane are maintained intact or alive. In a metaphorical sense, we can consider them as "living dead" or "zombie cells". Stabilization of apoptotic cells can be used for reliable detection and quantification of apoptosis in cultured cells and may allow a safer administration of apoptotic cells in clinical applications. Furthermore, it opens new avenues in the functional reconstruction of apoptotic cells for longer preservation.
细胞凋亡的特征是细胞成分的降解,但细胞膜仍然完整。凋亡微管网络 (AMN) 在细胞凋亡过程中组织形成,在细胞膜下形成皮质结构,维持细胞膜的完整性。凋亡细胞的另一个特征是产生大量的活性氧 (ROS),这对细胞可能是有害的。本研究旨在开发一种方法,使凋亡细胞稳定化,用于诊断和治疗应用。我们通过使用含有紫杉醇(微管稳定剂)、Zn(半胱氨酸天冬氨酸蛋白酶抑制剂)和辅酶 Q(脂抗氧化剂)的鸡尾酒,可以将 H460 凋亡细胞在细胞培养物中稳定至少 72 小时,防止继发性坏死。稳定的凋亡细胞保持许多凋亡细胞的特征,如存在凋亡微管、细胞膜完整性、低细胞内钙水平、细胞膜电位、PS 外翻和被吞噬的能力。稳定的凋亡细胞可以被认为是死亡的细胞,其中细胞皮质和细胞膜保持完整或存活。从比喻意义上讲,我们可以将它们视为“活死人”或“僵尸细胞”。凋亡细胞的稳定化可用于可靠地检测和定量培养细胞中的细胞凋亡,并可能允许在临床应用中更安全地施用凋亡细胞。此外,它为更长时间的凋亡细胞功能重建开辟了新途径。