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Durotaxis Index of 3T3 Fibroblast Cells Scales with Stiff-to-Soft Membrane Tension Polarity.

ABSTRACT: Cell durotaxis is an essential process in tissue development. Although the role of cytoskeleton in cell durotaxis has been widely studied, whether cell volume and membrane tension are implicated in cell durotaxis remains unclear. By quantifying the volume distribution during cell durotaxis, we show that the volume of 3T3 fibroblast cells decreases by almost 40% as cells migrate toward stiffer regions of gradient gels. Inhibiting ion transporters that can reduce the amplitude of cell volume decrease significantly suppresses cell durotaxis. However, from the correlation analysis, we find that durotaxis index does not correlate with the cell volume decrease. It scales with the membrane tension difference in the direction of stiffness gradient. Because of the tight coupling between cell volume and membrane tension, inhibition of Na⁺/K⁺ ATPase and Na⁺/H⁺ exchanger results in smaller volume decrease and membrane tension difference. Collectively, our findings indicate that the polarization of membrane tension is a central regulator of cell durotaxis, and Na⁺/K⁺ ATPase and Na⁺/H⁺ exchanger can help to maintain the membrane tension polarity.

SUBMITTER: Yang Y

PROVIDER: S-EPMC7567979 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Durotaxis, biased cell movement up a stiffness gradient on culture substrates, is one of the useful taxis behaviors for manipulating cell migration on engineered biomaterial surfaces. In this study, long-term durotaxis was investigated on gelatinous substrates containing a soft band of 20, 50, and 150 ?m in width fabricated using photolithographic elasticity patterning; sharp elasticity boundaries with a gradient strength of 300 kPa/50 ?m were achieved. Time-dependent migratory behaviors of 3T3 fibroblast cells were observed during a time period of 3 days. During the first day, most of the cells were strongly repelled by the soft band independent of bandwidth, exhibiting the typical durotaxis behavior. However, the repellency by the soft band diminished, and more cells crossed the soft band or exhibited other mixed migratory behaviors during the course of the observation. It was found that durotaxis strength is weakened on the substrate with the narrowest soft band and that adherent affinity-induced entrapment becomes apparent on the widest soft band with time. Factors, such as changes in surface topography, elasticity, and/or chemistry, likely contributing to the apparent diminishing durotaxis during the extended culture were examined. Immunofluorescence analysis indicated preferential collagen deposition onto the soft band, which is derived from secretion by fibroblast cells, resulting in the increasing contribution of haptotaxis toward the soft band over time. The deposited collagen did not affect surface topography or surface elasticity but did change surface chemistry, especially on the soft band. The observed time-dependent durotaxis behaviors are the result of the mixed mechanical and chemical cues. In the studies and applications of cell migratory behavior under a controlled stimulus, it is important to thoroughly examine other (hidden) compounding stimuli in order to be able to accurately interpret data and to design suitable biomaterials to manipulate cell migration.

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Durotaxis Index of 3T3 Fibroblast Cells Scales with Stiff-to-Soft Membrane Tension Polarity.

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