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Related Experiment Videos

Decrease in culture temperature releases monolayer endothelial cell sheets together with deposited fibronectin matrix

A Kushida1, M Yamato, C Konno

  • 1Institute of Biomedical Engineering, Tokyo Women's Medical University, Japan.

Journal of Biomedical Materials Research
|May 13, 1999
PubMed
Summary

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Researchers developed temperature-responsive polymer surfaces to culture endothelial cells. Lowering the temperature released intact cell sheets and associated fibronectin matrix, offering a novel approach for tissue engineering and extracellular matrix studies.

Area of Science:

  • Biomaterials Science
  • Cell Biology
  • Tissue Engineering

Background:

  • Cell culture typically requires enzymatic or mechanical dissociation for harvesting.
  • Maintaining cell-cell junctions and extracellular matrix integrity during cell recovery is challenging.
  • Temperature-responsive polymers offer potential for non-enzymatic cell detachment.

Purpose of the Study:

  • To evaluate poly(N-isopropylacrylamide) (PIPAAm) grafted surfaces for culturing bovine aortic endothelial cells.
  • To investigate the effect of temperature-responsive surfaces on cell adhesion, proliferation, and release.
  • To assess the recovery of intact cell sheets and associated fibronectin matrix.

Main Methods:

  • Culturing bovine aortic endothelial cells on PIPAAm-grafted surfaces.

Related Experiment Videos

  • Inducing cell release by lowering culture temperature.
  • Analyzing cell morphology and confluency.
  • Performing immunoblotting and immunofluorescence microscopy for fibronectin detection.
  • Comparing low-temperature release with enzymatic (trypsin) and mechanical methods.

    • Main Results:

    • Cells cultured on PIPAAm surfaces exhibited normal adhesion, spreading, proliferation, and confluency.
    • Lowering temperature induced non-enzymatic release of intact cell sheets from PIPAAm surfaces.
    • The PIPAAm surface transitioned from hydrophobic to hydrophilic upon cooling, weakening cell attachment.
    • Deposited fibronectin matrix was co-recovered with cell sheets upon temperature reduction.
    • Low-temperature recovery preserved the fibronectin matrix, unlike trypsin treatment.

    Conclusions:

    • Temperature-responsive PIPAAm surfaces enable non-enzymatic recovery of confluent endothelial cell sheets.
    • This method facilitates the retrieval of intact cell sheets and their associated extracellular matrix for tissue engineering.
    • The PIPAAm system provides a valuable tool for studying the dynamics of deposited extracellular matrix in cell culture.