细胞同步化技术在有丝分裂研究中的应用。

Cell Synchronization Techniques for Studying Mitosis.

机构信息

Department of Oncology, University of Alberta, Edmonton, AB, Canada.

Experimental Oncology, Cross Cancer Institute, Edmonton, AB, Canada.

出版信息

Methods Mol Biol. 2022;2579:73-86. doi: 10.1007/978-1-0716-2736-5_6.

DOI:10.1007/978-1-0716-2736-5_6

PMID:36045199

Abstract

Cell synchronization allows the examination of cell cycle progression. Nocodazole and other microtubule poisons have been used extensively to interfere with microtubule function and arrest cells in mitosis. Since microtubules are important for many cellular functions, alternative cell cycle synchronization techniques independent of microtubule inhibition are also used for synchronizing cells in mitosis. Here we describe using nocodazole, STLC, and combining thymidine block with MG132 to synchronize cells in mitosis. These inhibitors are reversible and mitotic cells can be released into the G1 phase synchronously. These techniques can be applied to both Western blot and timelapse imaging to study mitotic progression.

摘要

细胞同步化可以用来研究细胞周期的进程。诺考达唑和其他微管毒素已被广泛用于干扰微管功能，使细胞在有丝分裂中停滞。由于微管对于许多细胞功能都很重要，因此也有许多不依赖于微管抑制的细胞周期同步化技术用于使细胞同步化进入有丝分裂。在这里，我们描述了使用诺考达唑、STLC 以及将胸苷阻断与 MG132 相结合来使细胞同步化进入有丝分裂。这些抑制剂是可逆的，有丝分裂细胞可以同步释放到 G1 期。这些技术可以应用于 Western blot 和延时成像，以研究有丝分裂的进展。