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Related Experiment Videos

Temperature-dependent cell detachment on Pluronic gels.

Akon Higuchi1, Taro Yamamoto, Kaichiro Sugiyama

  • 1Department of Applied Chemistry, Seikei University, 3-3-1 Kichijoji Kitamachi, Musashino, Tokyo 180-8633, Japan. higuchi@ch.seikei.ac.jp

Biomacromolecules
|March 15, 2005
PubMed
Summary

Researchers developed a method using Pluronic gels for cell detachment at 4°C, enabling mild cell isolation. This technique enhances surface marker analysis for human umbilical vein endothelial cells (HUVECs) via flow cytometry.

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Area of Science:

  • Biomaterials Science
  • Cell Biology
  • Tissue Engineering

Background:

  • Pluronic gels, thermosensitive polymers, offer unique properties for cell culture.
  • Hydrophilicity of Pluronic gels can lead to dissolution in culture media.
  • Mild cell detachment methods are crucial for preserving cell surface markers.

Purpose of the Study:

  • To investigate temperature-dependent cell detachment from Pluronic gels.
  • To evaluate the impact of extracellular matrix (ECM) on cell detachment and surface marker expression.
  • To assess the utility of this method for surface marker analysis in human umbilical vein endothelial cells (HUVECs).

Main Methods:

  • Cell culture of HUVECs on Pluronic gels with and without ECM.
  • Temperature-induced cell detachment by cooling to 4°C.

Related Experiment Videos

  • Surface marker analysis (CD34, CD105) using flow cytometry.

    • Main Results:

    • Successful temperature-dependent cell detachment from Pluronic gels was achieved.
    • Cells detached from Pluronic gels lacking ECM showed a twofold higher CD34/CD105 expression ratio compared to trypsinized cells from polystyrene.
    • Higher ECM concentrations on gels reduced the observed expression ratios, potentially due to antibody binding interference.

    Conclusions:

    • Temperature-dependent cell detachment from Pluronic gels provides a mild isolation method.
    • This technique is a valuable tool for surface marker analysis, particularly for HUVECs.
    • Careful consideration of ECM concentration is necessary to avoid interference with antibody-based detection.