# Controlled Exit from the G2/M Checkpoint in RPE-1 Cells Using RO3306: Enrichment of Phase-Specific Cell Populations for In-Depth Analyses of Mitotic Events

## TL;DR

This paper introduces a new method to synchronize cells at specific mitotic stages using RO3306, enabling detailed study of cell division without disrupting normal processes.

## Contribution

A novel RO3306-based synchronization strategy that preserves mitotic integrity and allows high-resolution analysis of mitotic events.

## Key findings

- RO3306 block-and-release strategy enriches mitotic cell populations without disrupting mitotic progression.
- The method preserves cell viability and mitotic integrity, avoiding chromosome non-disjunction and other side effects.
- This approach provides a reliable framework for studying genomic instability and cell division mechanisms.

## Abstract

Studying the cell cycle is essential for understanding the molecular mechanisms that regulate cell division, growth, and differentiation in living organisms. However, mitosis constitutes only a brief phase of the overall cell cycle, making its analysis challenging in asynchronous cell populations due to its transient and dynamic nature. Cell synchronization methods help to enrich populations at specific cell cycle stages, including mitosis, typically by using chemical inhibitors to arrest cells at defined checkpoints. However, many existing protocols rely on combinations of inhibitors that interfere with normal mitotic progression, disrupting dynamics and causing side effects such as chromosome non-disjunction or lagging chromosomes, which limit their applicability. In this study, we present an RO3306 block-and-release strategy to selectively enrich cell populations at defined mitotic stages without compromising cell viability or disrupting their progression to mitotic exit. This approach provides a reliable method for studying mitotic events with high temporal resolution. Furthermore, by preserving mitotic integrity, it offers a valuable framework for investigating the molecular mechanisms of cell division and the processes driving genomic instability in human cells.