Cell Synchronization Techniques to Study the Action of CDK Inhibitors

Methods Mol Biol. 2016:1336:85-93. doi: 10.1007/978-1-4939-2926-9_8.

Abstract

Cell synchronization techniques have been used for the studies of mechanisms involved in cell cycle regulation. Synchronization involves the enrichment of subpopulations of cells in specific stages of the cell cycle. These subpopulations are then used to study regulatory mechanisms of the cell cycle such as DNA synthesis, gene expression, protein synthesis, protein phosphorylation, protein degradation, and development of new drugs (e.g., CDK inhibitors). Here, we describe several protocols for synchronization of cells from different phases of the cell cycle. We also describe protocols for determining cell viability and mitotic index and for validating the synchrony of the cells by flow cytometry.

MeSH terms

  • Animals
  • Aphidicolin / chemistry
  • Cell Culture Techniques / methods*
  • Cell Cycle
  • Cell Line, Tumor
  • Cell Proliferation
  • Cell Survival
  • Cyclin-Dependent Kinase Inhibitor Proteins / chemistry*
  • Cyclin-Dependent Kinases / antagonists & inhibitors*
  • DNA / chemistry
  • DNA Replication
  • Flow Cytometry
  • HeLa Cells
  • Humans
  • Mice
  • Mitosis
  • Mitotic Index
  • NIH 3T3 Cells
  • Nocodazole / chemistry
  • Protein Kinase Inhibitors / chemistry*
  • Thymidine / chemistry
  • Time Factors
  • Trypan Blue / chemistry

Substances

  • Cyclin-Dependent Kinase Inhibitor Proteins
  • Protein Kinase Inhibitors
  • Aphidicolin
  • DNA
  • Cyclin-Dependent Kinases
  • Trypan Blue
  • Nocodazole
  • Thymidine