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Tumor Cell Invasion Assay: A 3D In Vitro Native Matrix-based Method to Assess Epithelial-mesenchymal Cell Interactions
In This Article
Overview
This video describes the technique of utilizing a native matrix to study tumor cell invasion. The technique used in this assay provides the ability to assess epithelial-mesenchymal cell interactions in a 3D setting without the need for a synthetic or foreign matrix.
Protocol
1. Preparation of Media and Reagents
- Preparation of 200x of L-ascorbic acid 2-phosphate stock
- Dissolve 29 mg of L-ascorbic acid 2-phosphate per 5 mL of Dulbecco's Modified Eagle's Medium (DMEM) solution and filter through a 0.22 μm membrane filter. Store as 0.25 mL sterile aliquots at -20 °C.
- Add 0.25 mL aliquot of 200x L-ascorbic acid 2-phosphate stock to every 50 mL of fibroblast media (DMEM with 1% L-glutamine and 10% fetal bovine serum) on the day required, for a final concentration of 0.1 mM of L-ascorbic acid 2-phosphate.
- Preparation of keratinocyte growth media
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Representative Results
Figure 1. Workflow of generation of fibroblast-derived native matrix and the tumor invasion assay.
This article has been published
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Source: Ng, Y. Z. et al. Tissue Engineering of Tumor Stromal Microenvironment with Application to Cancer Cell Invasion. J. Vis. Exp. (2014)
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