Cell Adhesion Assay
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The document describes a cellular detachment assay using a parallel-plate flow chamber to study T lymphocyte adhesion under varied shear stress conditions. It details the components and settings of the assay, including the use of fibronectin and monoclonal antibodies for T cell adhesion measurements. Shear stress levels for both venous and arterial conditions are provided, referencing relevant literature.
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Cell Adhesion Assay
- 1. Cellular detachment assay Schematic for Parallel-plate Flow (PPF) Hard plastic chamber Plate with ECM absorbed Thin rubber gasket buffer buffer + cells
- 2. T lymphocytes adhered to fibronectin
- 3. Activated T lymphocytes adhered to fibronectin
- 4. Shear Stress 6 μ Q w h 2 µ = viscosity of the media (0.007) Q = the flow rate in cm 3 /s (dyne/cm 2 ) w = width of chamber h = height of chamber Venous shear stress: 1-6 dyne/cm 2 Arterial shear stress: 10-70 dyne/cm 2 (Malek, et al., JAMA, 1999)
- 5. Data: Activated T cell adhesion to FN
- 6. Data: CD45RA and RO T cell adhesion to monoclonal antibodies (dyne/cm 2 )