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To determine whether the difference in phagocytosis rates of white and opaque cells reflects a difference in adherence, we modified the assay for use at 4°C, a temperature that allows adherence but blocks phagocytosis (see Materials and Methods).
Binding of CXCR-4 to CXCL12 allows adherence and extravasation of OS cells in the lung [ 9, 10, 14, 15, 22, 42, 69].
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29 32 36 Our analysis differs from others in assuming that the therapies are identical in terms of clinical efficacy among adherent patients, but allowing adherence rates to determine differences in CV events and economic outcomes.
The in vitro cell binding was assessed by growing B16F1 cells in culture flasks to 80 90% confluence, trypsinizing, washing in PBS (pH 7.4), aliquoting into 12-well plates and allowing adherence to the wells overnight.
MEFs were seeded onto 6-well-plates and incubated overnight to allow adherence.
MEFs were seeded onto coverslips in 12-well-plates and incubated overnight at 37°C and 5% CO2 to allow adherence.
These fragments were placed on the surface of 100 mm Petri dishes for 40 50 minutes to allow adherence of the tissue to the culture surface.
Lymphocytes were enriched by incubation in complete medium (described above) in sterile tissue culture 10 cm plates (Falcon, USA) for overnight in 37 degrees Celsius to allow adherence of cells.
Thus, it is possible that Lyme disease borreliae capture CFHRs or lipoprotein particles to allow adherence to host epithelial cells and tissues, as recently described for CFH-coated S. pneumoniae [59].
Cell viability was again determined by trypan blue dye exclusion and if improved to >50% cells were plated out at 5×105 per well in supplemented media for 2 hours onto type 1 rat collagen coated plates to allow adherence of cells.
After incubation, the cells were washed twice by centrifugation and resuspension in AIP; ∼4·106 cells were then transferred to a chamber with glass bottom coated with poly-L-lysine (#1.0, MatTek Corp., Ashland, MA, USA) and further incubated at 37°C for 60 minutes to allow adherence to the coverslip.
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