Tumor Cell Invasion Assay: A 3D In Vitro Native Matrix-based Method to Assess Epithelial-mesenchymal Cell Interactions
Published: April 30, 2023
Abstract
Source: Ng, Y. Z. et al. Tissue Engineering of Tumor Stromal Microenvironment with Application to Cancer Cell Invasion. J. Vis. Exp. (2014)
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 …
Representative Results
Figure 1. Workflow of generation of fibroblast-derived native matrix and the tumor invasion assay.
Divulgazioni
The authors have nothing to disclose.
Materials
| L-Ascorbic acid 2-phosphate | Sigma | A8960 | |
| DMEM with L-glutamine, 4,500 mg/L D-glucose, 110 mg/L sodium pyruvate | Life Technologies | 11995-073 | |
| 100x Penicillin-Streptomycin | Life Technologies | 15070 | |
| Vaseline | VWR | PROL28908.290 | |
| Clonal cylinders | Sigma | Z370789 | |
| Nylon Net Filter Disc Hydrophilic 100 μm 25 μm diameter 100/pk | Millipore | NY1H02500 | |
| Bent stainless steel wire mesh support | Made in house | Dimensions were made so that the mesh would fit into 6-well plates |
Citazione di questo articolo
Tumor Cell Invasion Assay: A 3D In Vitro Native Matrix-based Method to Assess Epithelial-mesenchymal Cell Interactions. J. Vis. Exp. (Pending Publication), e20366, doi: (2023).