Microfluidic Shear Force Assay to Determine Cell Adhesion Forces

Methods Mol Biol. 2019:2017:71-84. doi: 10.1007/978-1-4939-9574-5_6.

Abstract

Cell adhesion is implicated in many physiological settings such as the retention of hematopoietic stem cells (HSCs) in their bone marrow niches or their migration into the bloodstream. During HSC mobilization these adhesion sites are cleaved and have to be newly formed during HSC homing and engraftment. To determine the adhesive properties of HSCs on different extracellular matrix (ECM) molecules, we present a microfluidic shear force assay, where a laminar flow is used to detach a semi-adherent cell population, the HSC model cell line KG-1a, from an ECM protein-coated substrate. This technique combines the high throughput of population-based assays with the ability to observe cell detachment in real time. Additionally, it is suitable for weakly adherent cells, as the setup allows cell incubation on various substrates and application of shear stress ranging several orders of magnitude in one setup without additional washing or transfer steps. As a measure for the adhesion strength of the studied cell population on the substrate, the critical shear force τ50 is determined which is required to remove 50% of the initially adherent cell fraction.

Keywords: Cell adhesion; Leukemic cells; Microfluidic assay; Protein coating; Shear force.

MeSH terms

  • Cell Adhesion
  • Cell Line
  • Cell Movement
  • Extracellular Matrix / metabolism
  • Hematopoietic Stem Cells / cytology*
  • Humans
  • Microfluidics / instrumentation*
  • Stress, Mechanical