Mesenchymal Stromal Cells as Active Regulators of Lymphocyte Recruitment to Blood Vascular Endothelial Cells

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)

Colleges, School and Institutes

External organisations

  • Institute of Cardiovascular Sciences, The Medical School, University of Birmingham, Birmingham, B15 2TT, UK.

Abstract

Methods are described for analyzing adhesion and migration of isolated lymphocytes on endothelial cell monolayers which have been cocultured with different mesenchymal stromal cells, with or without additional cytokine treatment. The different cells types are grown on opposite sides of 3.0 or 0.4 μm pore filters, depending on whether migration through the whole construct is to be analyzed, or adhesion to the endothelial cells alone. Migration away from the sub-endothelial space and through the stromal layer can also be assessed by culturing mesenchymal stromal cells within a 3-D collagen gel overlaid with endothelial cells. Assays may be "static" or the filter-based constructs can be incorporated into flow chambers so that cell behavior can be directly observed under conditions simulating those in vivo. In general, by choice of method, one can evaluate efficiency of attachment, and ability of cells to migrate across the endothelial monolayer, through the filter and through the stromal cell layer in 2-D or 3-D. Fluorescence microscopic examination of fixed filters can be used, e.g., to ascertain whether lymphocytes are retained by stromal cells. In general, static assays have the higher throughput and greatest ease of use, while the flow-based assays are more physiologically relevant and allow detailed recording of cell behavior in real time.

Details

Original languageEnglish
Title of host publicationT-Cell Trafficking
Subtitle of host publicationMethods and Protocols
EditorsGeorge Edward Rainger, Helen M. Mcgettrick
Publication statusE-pub ahead of print - 28 Mar 2017

Publication series

NameMethods in Molecular Biology
PublisherSpringer New York
Volume1591
ISSN (Electronic)1064-3745

Keywords

  • Lymphocyte, Endothelial cells, Fibroblasts, Mesenchymal stem cells MSC , Stromal cells, Adhesion, Migration, Cytokines, Cell culture, Coculture, Inflammation