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Altered lipid packing identifies apoptotic thymocytes

R A Schlegel1, M Stevens, K Lumley-Sapanski

  • 1Department of Molecular and Cell Biology, Pennsylvania State University, University Park 16802.

Immunology Letters
|June 1, 1993
PubMed
Summary
This summary is machine-generated.

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Researchers identified early apoptotic thymocytes using merocyanine 540 (MC540) staining, which detects changes in plasma membrane lipid packing. This method allows for the separation of viable apoptotic cells based on their altered membrane properties.

Area of Science:

  • Immunology
  • Cell Biology
  • Biophysics

Background:

  • Apoptosis is a critical process in thymocyte development.
  • Identifying apoptotic cells in real-time is crucial for understanding immune regulation.
  • Current methods for detecting apoptosis may not capture early, viable stages.

Purpose of the Study:

  • To investigate if altered plasma membrane lipid packing in apoptotic thymocytes can be detected.
  • To determine if merocyanine 540 (MC540) can identify these membrane changes.
  • To establish a method for isolating viable apoptotic thymocytes.

Main Methods:

  • Thymocytes were isolated from hydrocortisone-injected mice.
  • Cells were stained with merocyanine 540 (MC540), a lipid packing-sensitive fluorescent probe.

Related Experiment Videos

  • DNA degradation was assessed using propidium iodide (PI) staining.
  • Fluorescence-activated cell sorting (FACS) was used for cell enrichment.

    • Main Results:

    • A subpopulation of H2-Klo cells with increased MC540 staining emerged 9-10 hours post-injection.
    • Increased MC540 staining correlated with reduced PI staining, indicating DNA degradation.
    • FACS enrichment of MC540-high cells also enriched for PI-low cells.
    • Living apoptotic thymocytes exhibit altered lipid packing and increased MC540 uptake.

    Conclusions:

    • Merocyanine 540 (MC540) is a reliable marker for identifying early apoptotic thymocytes.
    • Altered plasma membrane lipid packing is an early event in thymocyte apoptosis.
    • MC540 staining combined with cell sorting enables the isolation of viable apoptotic thymocytes.