A methodology to study chemotaxis in 3-D collagen gels

Sergio Caserta, Silvia Campello, Giovanna Tomaiuolo, Luigi Sabetta, Stefano Guido

Research output: Contribution to journalArticlepeer-review

Abstract

We present here an innovative experimental methodology for the quantitative investigation of chemotaxis in vitro by live imaging of cell movement in a reconstituted three-dimensional collagen gel. A well-defined chemoattractant gradient is generated by means of a novel direct viewing chamber having two compartments (separated by a membrane), one containing the chemoattractant solution, the other the cell-seeded collagen gel matrix. Cell migration is observed by means of a time-lapse motorized video-microscopy workstation equipped with an incubating system and quantified by image analysis techniques. Experimental results on three different cell lines (Jurkat, fibroblasts, and lymphocytes) are presented for the isotropic control case (no chemoattractant) and in presence of a concentration gradient. Cell motility data are in line with the concentration profile, both theoretically calculated from Fick's law and experimentally measured by epifluorescence microscopy. In particular, a transient peak in cell response was found, possibly due to cell membrane receptor saturation.

Original languageEnglish
Pages (from-to)4025-4035
Number of pages11
JournalAICHE Journal
Volume59
Issue number11
DOIs
Publication statusPublished - Nov 2013

Keywords

  • 3-D cell tracking
  • Cell motility
  • Chemotaxis
  • Collagen gel
  • Diffusion
  • Time-lapse microscopy

ASJC Scopus subject areas

  • Chemical Engineering(all)
  • Biotechnology
  • Environmental Engineering

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