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Stripe Assay to Study the Attractive or Repulsive Activity of a Protein Substrate Using Dissociated Hippocampal Neurons

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Published on: June 19, 2016

Combinatorial nanodot stripe assay to systematically study cell haptotaxis.

Mcolisi Dlamini^1,2,3, Timothy E Kennedy^3,4, David Juncker^1,2,3,4

¹Biomedical Engineering Department, McGill University, 3775 University Street, Montréal, QC H3A 2B4 Canada.

Microsystems & Nanoengineering

|December 28, 2020

Summary

This summary is machine-generated.

New nanodot stripe assays (NSAs) allow precise study of cell haptotaxis. Cells showed a clear preference for higher netrin-1 surface coverage, revealing new insights into cell guidance mechanisms.

Keywords:

Nanofabrication and nanopatterning Nanoscale materials

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Area of Science:

Cell Biology
Biophysics
Materials Science

Background:

Haptotaxis, or cell movement along substrate-bound cues, is vital for development.
Existing in vitro methods like gradients and stripe assays have limitations in studying haptotaxis.
These limitations include receptor saturation, migration history, and binary choices.

Purpose of the Study:

To develop a novel assay for quantitative analysis of cell haptotaxis across a wide range of surface coverages.
To investigate the migration behavior of C2C12 myoblasts on patterned nanodot surfaces with varying netrin-1 concentrations.
To elucidate the quantitative relationship between surface cue density and cell migration preference.

Main Methods:

Introduction of nanodot stripe assays (NSAs) with adjacent stripes of varying nanodot surface coverages (0-100%).
Patterning of nanodots in sizes 200x200, 400x400, or 800x800 nm² on surfaces.
Studying C2C12 myoblast migration on netrin-1 patterned NSAs and three-step gradients.

Main Results:

Cell response to haptotaxis was independent of nanodot size.
Cells increasingly preferred higher netrin-1 coverage stripes, with up to 90% preference at 10% coverage and higher.
A preference for higher netrin-1 coverage was observed only for coverage ratios greater than 2.3, plateauing around 80% for ratios >= 4.

Conclusions:

The combinatorial NSA provides a powerful tool for quantitative cell haptotaxis studies over the full spectrum of surface coverages.
This method allows for a more nuanced understanding of haptotactic mechanisms than previously possible.
Findings reveal specific thresholds and saturation points in cell preference for cue density.