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Cell Blebbing in Confined Microfluidic Environments.

Markela Ibo¹, Vasudha Srivastava², Douglas N Robinson²

¹Johns Hopkins University, Department of Chemical and Biomolecular Engineering, Baltimore, MD, 21218, United States of America.

|October 6, 2016

Summary

This summary is machine-generated.

Dictyostelium cells switch to bleb-based migration in confined environments. Increased cAMP gradient steepness and reduced channel height promote bleb formation by influencing Myosin II activity and intracellular pressure.

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Area of Science:

Cell Biology
Biophysics

Background:

Cell migration involves protrusions like blebs and pseudopods.
Dictyostelium discoideum cells alter migration strategy in resistive environments, favoring blebs.

Purpose of the Study:

Investigate Dictyostelium discoideum blebbing responses to cAMP gradients in microfluidic channels.
Determine the influence of microfluidic confinement height and gradient steepness on cell migration modes.

Main Methods:

Utilized microfluidic channels with varying confinement heights (1.7–3.8 μm) to create cAMP gradients.
Observed and analyzed Dictyostelium cell migration patterns, bleb formation, and pseudopod dynamics.
Manipulated buffer osmolarity and Myosin II activity (using blebbistatin) to assess their effects.

Main Results:

Cells shifted from mixed bleb/pseudopod migration to exclusive bleb migration as cAMP gradient steepness increased (0.7 to 20 nM/μm) in channels < 2.4 μm height.
Bleb size increased with gradient steepness, correlating with enhanced Myosin II localization.
Reduced intracellular pressure (high osmolarity) or inhibited Myosin II decreased bleb formation and size.

Conclusions:

Microfluidic confinement height and cAMP gradient steepness significantly influence Dictyostelium cell protrusion type.
Bleb formation is regulated by a combination of confinement-induced Myosin II localization and cAMP-driven cortical contraction, increasing intracellular pressure.