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Related Experiment Videos

In vitro neutrophil transepithelial migration.

Winston Y Lee1, Alex C Chin, Susan Voss

  • 1Division of Gastrointestinal Pathology, Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 27, 2006
PubMed
Summary

This study presents a new in vitro model to investigate polymorphonuclear leukocyte (PMN) migration across intestinal epithelial cells. This method aids in understanding inflammatory diseases like ulcerative colitis and Crohn's disease.

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Area of Science:

  • Immunology
  • Gastroenterology
  • Cell Biology

Background:

  • Polymorphonuclear leukocyte (PMN) transmigration is crucial for host defense but contributes to epithelial pathophysiology in inflammatory diseases.
  • The precise mechanisms regulating PMN migration across intestinal epithelium are not fully understood.

Purpose of the Study:

  • To describe a novel in vitro method for studying PMN transmigration across intestinal epithelial T84 monolayers.
  • To provide a reductionist and manipulable model system for investigating PMN responses in inflammatory conditions.

Main Methods:

  • Utilized a modified Boyden chamber system with inverted T84 intestinal epithelial monolayers on permeable supports.
  • Isolated human PMNs from blood via dextran sedimentation and Ficoll density gradient centrifugation.

Related Experiment Videos

  • Elicited PMN transmigration using N-formyl-methionyl-leucyl-phenylalanine gradients and quantified via myeloperoxidase activity.

    • Main Results:

    • Successfully established an in vitro model for studying PMN transmigration in a physiologically relevant basolateral-to-apical direction.
    • The model allows for controlled investigation of PMN migration mechanisms.
    • Quantification of transmigration was achieved through myeloperoxidase activity assays.

    Conclusions:

    • The developed in vitro model offers a valuable tool for dissecting the complex mechanisms of PMN transmigration.
    • This research will advance our understanding of PMN roles in acute inflammatory diseases affecting the intestine.