Kato Takamitsu A
Department of Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA.
Methods Mol Biol. 2025;2933:81-85. doi: 10.1007/978-1-0716-4574-1_11.
The mitotic shake-off method is a laboratory technique used to isolate cells that are actively undergoing mitosis from a population of cells in culture. During mitosis, cells undergo a series of characteristic changes. After the cells have been induced to enter mitosis, they are gently dislodged from the culture vessel using a mild mechanical agitation technique, such as shaking or tapping. Since cells in mitosis typically round up and detach from the substrate as they prepare to divide, they are more easily released from the culture surface compared to cells in other phases of the cell cycle. Those isolated mitotic cells can be transferred to other cell culture vessels to release cell cycle progression. Mitotic cells go through next cell cycle phases with time. Using this method, cell cycle-dependent cellular activity such as cell cycle-specific protein expression and activation, and cell cycle-specific sensitivity to agents, can be carried out. Since this method is completely drug free, it has series of benefits including no drug cost and no potential toxicity from drug exposure. This chapter introduces cellular synchronization technique by mitotic shake-off method.
有丝分裂抖落法是一种实验室技术，用于从培养的细胞群体中分离出正在进行有丝分裂的活跃细胞。在有丝分裂过程中，细胞会经历一系列特征性变化。在细胞被诱导进入有丝分裂后，使用温和的机械搅拌技术，如摇晃或轻敲，将它们从培养容器中轻轻移出。由于处于有丝分裂期的细胞在准备分裂时通常会变圆并从底物上脱离，与细胞周期其他阶段的细胞相比，它们更容易从培养表面释放出来。那些分离出的有丝分裂细胞可以转移到其他细胞培养容器中以释放细胞周期进程。随着时间的推移，有丝分裂细胞会经历下一个细胞周期阶段。使用这种方法，可以进行细胞周期依赖性细胞活动，如细胞周期特异性蛋白质表达和激活，以及细胞周期特异性对试剂的敏感性。由于该方法完全无药物，它具有一系列优点，包括无药物成本和无药物暴露潜在毒性。本章介绍通过有丝分裂抖落法进行细胞同步化技术。
