Send to

Choose Destination
J Vis Exp. 2019 May 24;(147). doi: 10.3791/59226.

3D Analysis of Multi-cellular Responses to Chemoattractant Gradients.

Author information

Department of Biomedical Engineering and Yale Systems Biology Institute, Yale University.
Department of Biomedical Engineering, Johns Hopkins University.
Department of Biomedical Engineering, Johns Hopkins University; Center for Cell Dynamics and Department of Cell Biology, Johns Hopkins University.
Department of Biomedical Engineering and Yale Systems Biology Institute, Yale University;


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.


Supplemental Content

Full text links

Icon for MyJove Corporation
Loading ...
Support Center