Optimizing Cell Synchronization Using Nocodazole or Double Thymidine Block

Methods Mol Biol. 2021:2329:111-121. doi: 10.1007/978-1-0716-1538-6_9.

Abstract

Cell synchronization is crucial when studying events that take place at specific points of the cell cycle. Several chemical agents can be used to achieve the cell culture synchronization but not all type of cells respond equally to a given concentration of these drugs. Here we describe a simple optimization method to select concentrations and timings for nocodazole or thymidine treatments using fluorescence staining. In addition, we provide detailed protocols to arrest an asynchronous culture of either suspension or adherent cells in G1/S or in G2/M.

Keywords: Cell cycle; G1/S; G2/M; Nocodazole; Suspension cells; Synchronization; Thymidine.

MeSH terms

  • Cell Adhesion
  • Cell Culture Techniques / methods*
  • Cell Cycle
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Dose-Response Relationship, Drug
  • Fluorescent Dyes / chemistry
  • HeLa Cells
  • Humans
  • Nocodazole / pharmacology*
  • Thymidine / pharmacology*
  • Time Factors

Substances

  • Fluorescent Dyes
  • Nocodazole
  • Thymidine