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Cell migration is a process by which the cells move from one location to another, playing an essential role in embryological development, repair and regeneration, immune response, and metastasis. Cells migrate in response to chemical or mechanical signals generated by specific organs or tissues. The overall mechanism includes three steps - polarization, protrusion, and release. Polarization involves the formation of a distinct cell front and rear, which determines the direction of movement.
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Cells can detect chemical cues in their environment and reorganize the cytoskeleton to migrate toward them or away from them. This directional migration, called chemotaxis, is essential during embryogenesis and development, immune response, tissue repair and regeneration, and reproduction. These chemical cues can either attract or repel the cell's movement. For example, axon development is determined by a combination of chemoattractants and chemorepellents that direct the growing axon...
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Live-Cell Imaging Assays to Study Glioblastoma Brain Tumor Stem Cell Migration and Invasion
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An optimized method for accurate quantification of cell migration using human small intestine cells.

Steffen Nyegaard1, Brian Christensen1, Jan Trige Rasmussen1

  • 1Department of Molecular Biology and Genetics - Molecular Nutrition, Aarhus University, Gustav Wieds Vej 10, DK-8000 Aarhus C, Denmark.

Metabolic Engineering Communications
|February 23, 2018
PubMed
Summary

A new Cell Exclusion Zone Assay offers a reliable method to quantify cell migration, overcoming limitations of traditional scratch assays. This technique accurately measures how compounds affect cell movement, crucial for understanding diseases like cancer.

Keywords:
(BME), Basal membrane extract(Caco-2), human epithelial colorectal adenocarcinoma cells(DMEM), Dulbecco's modified Eagle medium(ECM), Extracellular matrix(EGF), Recombinant human epidermal growth factor(FBS), fetal bovine serum(FHs-74 int), non-malignant human fetal small intestine cells(FRET), Förster resonance energy transfer(OPN), osteopontin(ROI), region of interestBioactiveCollective migrationEpitheliumMigration assaySmall intestine cellsWound healing

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Three-dimensional Quantification of Intestinal Mucus Using Whole-mount Tissue Imaging
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Area of Science:

  • Cell Biology
  • Biotechnology
  • Biophysics

Background:

  • Quantifying cell migration is vital for research in chemotaxis, wound healing, and cancer metastasis.
  • Traditional scratch assays have limitations, including potential cell stress and damage to underlying matrices, which can skew results.
  • A novel Cell Exclusion Zone Assay (CEZA) is presented to address these limitations.

Purpose of the Study:

  • To develop and validate a new assay for quantifying cell migration rates.
  • To assess the impact of specific substrates and bioactive proteins on human small intestine cell migration.
  • To establish a reliable and versatile method for high-throughput cell migration analysis.

Main Methods:

  • The Cell Exclusion Zone Assay involves creating cell-free areas using removable silicone stoppers.
  • Human small intestine epithelial cells were cultured on various substrates to optimize adherence and migration.
  • Cell migration was quantified by counting nuclei that entered the cell-free zone after fluorescent staining.

Main Results:

  • The CEZA successfully quantified cell migration on different substrates, monitoring morphological changes.
  • Recombinant human epidermal growth factor and osteopontin demonstrated dose-dependent modulation of human small intestine cell migration.
  • The assay proved accurate and reliable for assessing the effects of bioactive proteins on cellular migration.

Conclusions:

  • The Cell Exclusion Zone Assay provides a robust, versatile, and cost-effective alternative to traditional scratch assays.
  • This method is adaptable for various cell lines and matrices, enabling high-throughput analysis.
  • The assay facilitates intricate analysis of mixed cell populations and can be combined with co-staining for comprehensive cell behavior studies.