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inflammation - Inflammation Chapter 2 Chemotaxis and...

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Inflammation Chapter 2 Feb 16, 2005
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Chemotaxis and chemoinvasion assays Migration and invasion was assayed in 24-well cell-culture chambers using inserts with 8-μm pore membranes as described. Membranes were pre-coated with fibronectin (2.5–7.5 μg ml- 1) for chemotaxis or Matrigel (28 μg per insert) and fibronectin for invasion studies. Breast cancer cells were resuspended in chemotaxis buffer (DMEM/ 0.1% BSA/ 12 mM HEPES) at 2 or 4 10 4 cells per ml. After incubation for 6 or 24 h for chemotaxis or chemoinvasion assays, respectively, cells on the lower surface of the membrane were stained and counted under a light microscope in at least five different fields (original magnification, 200). Assays were performed in triplicates. Chemokinesis was tested in checkerboard assays and was uniformly negative for both CXCL12 and CCL21.
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Boyden chamber Figures removed for copyright reasons.
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Checkerboard assay nM Top Bottom 0 1 10 100 0 0.2 0.2 0.3 0.5 1 0.8 0.3 0.4 0.4 10 2.7 2.3 0.6 0.6 100 3.4 3.5 0.8 0.7
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Figures removed for copyright reasons.
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Cells Behave Better on BD Matrigel™ Matrix BD Matrigel™ Matrix is a solubulized basement membrane preparation extracted from EHS mouse sarcoma, a tumor rich in ECM proteins. Its major component is laminin, followed by collagen IV, heparan sulfate proteoglycans, and entactin.
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