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Mass Cytometric Cell Cycle Analysis.

Gregory K Behbehani1

  • 1Division of Hematology, The Ohio State University and James Cancer Hospital, Columbus, OH, USA. Gregory.behbehani@osumc.edu.

Methods in Molecular Biology (Clifton, N.J.)
|May 12, 2019
PubMed
Summary
This summary is machine-generated.

Mass cytometry offers a powerful method to analyze cell cycle progression alongside numerous cellular markers. This technique enables detailed characterization of cell cycle interactions in various biological samples.

Keywords:
Cell cycleCyTOFCyclinIodo-deoxyuridineKi-67Mass cytometryPhosphorylated Histone H3Retinoblastoma protein

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Area of Science:

  • Cell Biology
  • Immunology
  • Oncology

Background:

  • Regulated cell proliferation is fundamental to multicellular organism development, immune function, and cancer progression.
  • Traditional fluorescent flow cytometry for cell cycle analysis has limitations in multiplexing capabilities.
  • Mass cytometry provides an advanced platform for simultaneous measurement of cell cycle and other cellular antigens.

Purpose of the Study:

  • To present a mass cytometry-based method for comprehensive cell cycle analysis.
  • To demonstrate the application of this method across diverse biological sample types.
  • To enable deeper understanding of cell cycle regulation in complex biological processes.

Main Methods:

  • Utilizing 5-Iodo-2'-deoxyuridine (IdU) incorporation for cell cycle phase labeling.
  • Measuring key cell cycle markers: phosphorylated retinoblastoma protein (pRb), Cyclin B1, and phosphorylated Histone H3 (pHH3).
  • Integrating these measurements into a robust gating strategy for precise cell cycle phase separation.

Main Results:

  • Successful application of the mass cytometry method to analyze cell cycle in vitro cultured cell lines.
  • Demonstrated efficacy in ex vivo analysis of human blood and bone marrow samples.
  • Validated the method for in vivo studies using murine tissues, achieving clear separation of all five cell cycle phases.

Conclusions:

  • Mass cytometry provides a versatile and high-dimensional approach for cell cycle analysis.
  • This method enhances the characterization of cell cycle dynamics in conjunction with other cellular processes.
  • The described technique is applicable across various research settings, from cell lines to complex tissues.