Laboratory of Physiological Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
Methods Mol Biol. 2025;2872:221-231. doi: 10.1007/978-1-0716-4224-5_15.
Centrosomes, the major microtubule organizing centers, facilitate mitotic spindle formation. However, recent studies have revealed that some cancer cells lack centrosomes. These findings suggest that certain types of cancer cells drive centrosome-independent mechanisms for the assembly of mitotic spindles. Therefore, analyzing the systems of acentrosomal spindle formation is beneficial for understanding the divergence of mitotic processes in cancer cells. Here, we focus on a method for generating acentrosomal cells using the PLK4 inhibitor centrinone and describe the proper conditions, spindle pole staining markers, and microscopy settings for observing acentrosomal spindle formation in human cells.
中心体是主要的微管组织中心，有助于有丝分裂纺锤体的形成。然而，最近的研究表明，一些癌细胞缺乏中心体。这些发现表明，某些类型的癌细胞驱动着中心体独立的机制来组装有丝分裂纺锤体。因此，分析无中心体纺锤体形成的系统有助于理解癌细胞中有丝分裂过程的分歧。在这里，我们重点介绍一种使用 PLK4 抑制剂 centrinone 产生无中心体细胞的方法，并描述了在人类细胞中观察无中心体纺锤体形成的适当条件、纺锤体极染色标记物和显微镜设置。
