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Cell Proliferation High-Content Screening on Adherent Cell Cultures.

Pau Carrillo-Barberà1,2,3, Jose M Morante-Redolat1,2,3, José F Pertusa4

  • 1Departamento de Biología Celular, Biología Funcional y Antropología Física, Universitat de València, Burjassot, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|August 22, 2019
PubMed
Summary
This summary is machine-generated.

This study presents a high-content proliferation assay for adherent cells using nucleoside analogues like EdU. The method quantifies cell cycle phases (G0-M) and proliferation rates in specific cell subpopulations.

Keywords:
Bioimage analysisCell proliferationEx vivoHigh-contentNeural stem cells

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biotechnology

Background:

  • Cell proliferation is crucial for development and disease.
  • Accurate measurement of cell cycle progression is essential for biological research.
  • Existing methods may lack the throughput or specificity required for complex analyses.

Purpose of the Study:

  • To describe a high-content proliferation assay pipeline for adherent cell cultures.
  • To enable detailed analysis of cell cycle phases and proliferation rates.
  • To facilitate the identification of specific cell subpopulations based on nuclear markers.

Main Methods:

  • Utilized pulse-chase experiments with nucleoside analogues (BrdU/EdU) to label cells in S phase.
  • Developed a high-throughput imaging and non-supervised ImageJ macroinstruction for data analysis.
  • Integrated nuclear segmentation, nucleoside analogue detection, and additional nuclear marker signal measurement.

Main Results:

  • Successfully quantified the percentage of cells in different cell cycle phases (G0, G1, S, G2, M).
  • Enabled estimation of proliferation (S phase) rates within specific cell subpopulations.
  • Demonstrated the pipeline's capability for high-content analysis of cell proliferation.

Conclusions:

  • The described pipeline offers a robust method for high-content proliferation analysis in adherent cells.
  • This assay allows for precise determination of cell cycle distribution and proliferation rates.
  • The method is adaptable for studying proliferation in distinct cell subpopulations identified by nuclear markers.