Study of cell deformability by a simple method |
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Authors: | Jean Louis Mege Christian Capo Anne Marie Benoliel Colette Foa Pierre Bongrand |
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Affiliation: | 1. Laboratoire d''Immunologie, Hôpital de Sainte Marguerite, B.P. 29, 13277 Marseille Cedex 09, France;2. Unité INSERM 119, 27 Bd. Leï Roure, 13009 Marseille, France |
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Abstract: | Cell deformability plays an important role in many immunological processes, such as phagocyte chemotaxis and endocytosis. The most widely used method of assay consists in aspirating cells into glass micropipettes and measuring the length of the protrusion induced by a given pressure, or the minimum pressure required to drive cells into the micropipette. This procedure requires specialized equipment and delicate manipulation. The present report describes a simpler procedure: cells are centrifuged in petri dishes floating on a water cushion, then fixed and coated with 0.8 μm diameter latex beads, which allows rapid and accurate determination of their height. This method is compared with the micropipette technique by studying lymphocyte and macrophage-like cell lines in physiological medium and in the presence of a divalent cation chelator or a microfilament inhibitor. In addition to simplicity, the main advantages of this technique are that (i) many cells may be examined within a reasonable period of time, which allows testing of heterogenous cell populations, and (ii) unexpectedly, centrifugation was quite harmless under our experimental conditions, since it did not impair cell proliferative ability nor phagocytic ability.It is concluded that the method may be used in clinical laboratories to explore phagocyte dysfunctions, as well as in experimental studies. |
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Keywords: | cell deformability cell spreading EGTA ethyleneglycol-bis aminoethyl ether tetraacetic acid ConA concanavalin A |
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