3D Analysis of Multi-cellular Responses to Chemoattractant Gradients
| Autor: | Sung Hoon Lee, Tae Yun Kang, Andre Levchenko, Andrew J. Ewald, David D. Ellison |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
General Immunology and Microbiology
Chemistry General Chemical Engineering General Neuroscience Context (language use) Matrix (biology) Organ-on-a-chip Receptors Formyl Peptide General Biochemistry Genetics and Molecular Biology Cell biology Extracellular Matrix Extracellular matrix 03 medical and health sciences Multicellular organism 3D cell culture Mice 0302 clinical medicine Imaging Three-Dimensional Cell culture 030220 oncology & carcinogenesis Organoid Animals Humans |
| Zdroj: | Journal of visualized experiments : JoVE. (147) |
| ISSN: | 1940-087X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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