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Biophysical Journal 86:1794-1806 (2004)
© 2004 The Biophysical Society

Nanometer Analysis of Cell Spreading on Matrix-Coated Surfaces Reveals Two Distinct Cell States and STEPs

Benjamin J. Dubin-Thaler, Gregory Giannone, Hans-Günther Döbereiner and Michael P. Sheetz

Department of Biological Sciences, Columbia University, New York, New York

Correspondence: Address reprint requests to Michael P. Sheetz, PhD, Dept. of Biological Sciences, PO Box 2408, Columbia University, Sherman Fairchild Center, Rm. 713, 1212 Amsterdam Ave., New York, NY, 10027. Tel.: 212-854-4857; Fax: 212-854-6399; E-mail: ms2001{at}columbia.edu.

When mouse embryonic fibroblasts in suspension contact a matrix-coated surface, they rapidly adhere and spread. Using total internal reflection fluorescence microscopy of dye-loaded fibroblasts to quantify cell-substrate contact, we found that increasing the surface matrix density resulted in faster spreading initiation whereas lamellipodial dynamics during spreading were unaltered. After spreading initiation, most cells spread in an anisotropic manner through stochastic, transient extension periods (STEPs) with ~30 STEPs over 10 min to reach an area of 1300 µm2 ± 300 µm2. A second mode of spreading, increased in serum-deprived cells, lacked STEPs and spread in a rapid, isotropic manner for 1–4 min. This isotropic mode was characterized by a high rate of area increase, 340 µm2/min with 78% of the cell edge extending. Anisotropic cells spread slower via STEPs, 126 µm2/min with 34% of the edge extending. During the initial 2–4 min of fast, isotropic spreading, centripetal flow of actin was low (0.8 µm/min) whereas in anisotropic cells it was high from early times (4.7 µm/min). After initial isotropic spreading, rearward actin movement increased and isotropic cells displayed STEPs similar to anisotropic cells. Thus, the two cell states display dramatically different spreading whereas long-term motility is based on STEPs.




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