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Force of cell adhesion after blocking CD11b or CD11c with antibodies.

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posted on 22.09.2016, 17:57 by Noémi Sándor, Szilvia Lukácsi, Rita Ungai-Salánki, Norbert Orgován, Bálint Szabó, Róbert Horváth, Anna Erdei, Zsuzsa Bajtay

MDDCs (A), MDMs (B) and monocytes (C) were treated with monoclonal anti-CD11b or anti-CD11c antibodies on ice for 30 min or left untreated for control. The Fc receptor blocking agent was used prior adding the antibodies in all samples. Cells were let to adhere for 30min at 37°C 5%CO2 in Petri dishes coated previously with 10μg/ml fibrinogen and blocked with PLL-g-PEG. After that cells were gently washed twice with PBS to remove unbound cells. The number of adhered cells was determined in the field of the microscope and is shown as 0,00μN. The computer controlled micropipette made serial pick-up processes in the field by using increasing amount of vacuum. A microscopic picture was taken after each round and the number of remaining cells was determined. The ratio of adhered cells was determined at each lifting force value by dividing the number of adhered cells in the anti-CD11b or anti-CD11c blocked samples by the number of cells in corresponding control sample. Data presented are mean +/-SD of three independent donors’ samples. Repeated measures ANOVA with Bonferroni post-test was used to determine significant differences compared to control at each force. * = p<0,05, ** = p<0,01.