Cell suspensions from monolayer cultures of 3T3, SV-3T3, BHK, BS-C-1, or macrophages, were prepared by brief exposure of the cultures to the amide anesthetic lidocaine. Cells were subcultivated six times without marked impairment of plating efficiency. The method should be applicable to physiological, biochemical, or immunological studies in which exposure of cells to proteolytic enzymes is to be avoided.

Cell functions such as adhesion, movement, and phagocytosis involve changes in both cell shape and cell surface motility. These changes are the result of membrane perturbations caused by interactions of cells with substrates, particles, or other cells. Surface perturbations generate signals that are presumably transduced into changes in the state of aggregation and distribution of submembranous cytoskeletal constituents (1-3). Except for the role of Fc and C3 receptors in the interaction of particles or substrates with phagocytes (3), other cell surface-"sensing" components, as well as the nature of the membrane perturbations and the transducing mechanisms are unknown. Induced cell shape changes permit the analysis of factors involved in maintenance of cell shape and in cell motility. A few diverse agents promote rapid and reversible changes in cell shape. Examples are the spreading of cells induced by Mn 2÷ (4) and the cell rounding induced by high hydrostatic pressure or exposure to trypsin, strychnine, EDTA, or cytochalasins (5-9). Cationic anesthetics inhibit a variety of cell surface phenomena, including cell to substrate adhesion, spreading, phagocytesis, locomotion, cell fusion, capping of surface Ig, or cell mediated-cytotoxicity (10-16). We show here that exposure of cultivated macrophages to cationic anesthetics results in extensive and fully reversible contraction and rounding. Single cells were followed before, during, and after exposure to the drugs and the observations recorded in still pictures and time-lapse movies. We examined several features of anesthetic-induced rounding, investigated structure-activity relationships of the drugs, and compared anesthetic effects to those of EDTA. Materials and Methods

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