3D Analysis of Multi-cellular Responses to Chemoattractant Gradients

Tae Yun Kang, David Ellison, Sung Hoon Lee, Andrew Ewald, Andre Levchenko

Research output: Contribution to journalArticle

Abstract

Various limitations of 2D cell culture systems have sparked interest in 3D cell culture and analysis platforms, which would better mimic the spatial and chemical complexity of living tissues and mimic in vivo tissue functions. Recent advances in microfabrication technologies have facilitated the development of 3D in vitro environments in which cells can be integrated into a well-defined extracellular matrix (ECM) and a defined set of soluble or matrix associated biomolecules. However, technological barriers have limited their widespread use in research laboratories. Here, we describe a method to construct simple devices for 3D culture and experimentation with cells and multicellular organoids in 3D microenvironments with a defined chemoattractant gradient. We illustrate the use of this platform for analysis of the response of epithelial cells and organoids to gradients of growth factors, such as epidermal growth factor (EGF). EGF gradients were stable in the devices for several days leading to directed branch formation in breast organoids. This analysis allowed us to conclude that collective gradient sensing by groups of cells is more sensitive vs. single cells. We also describe the fabrication method, which does not require photolithography facilities nor advanced soft lithography techniques. This method will be helpful to study 3D cellular behaviors in the context of the analysis of development and pathological states, including cancer.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number147
DOIs
StatePublished - May 24 2019

Fingerprint

Chemotactic Factors
Organoids
Cell culture
Epidermal Growth Factor
Tissue
Microfabrication
Biomolecules
Photolithography
Research laboratories
Lithography
Intercellular Signaling Peptides and Proteins
Cell Culture Techniques
Microtechnology
Fabrication
Equipment and Supplies
Extracellular Matrix
Breast
Epithelial Cells
Technology
Research

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

3D Analysis of Multi-cellular Responses to Chemoattractant Gradients. / Kang, Tae Yun; Ellison, David; Lee, Sung Hoon; Ewald, Andrew; Levchenko, Andre.

In: Journal of visualized experiments : JoVE, No. 147, 24.05.2019.

Research output: Contribution to journalArticle

Kang, Tae Yun ; Ellison, David ; Lee, Sung Hoon ; Ewald, Andrew ; Levchenko, Andre. / 3D Analysis of Multi-cellular Responses to Chemoattractant Gradients. In: Journal of visualized experiments : JoVE. 2019 ; No. 147.
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