I am working on a cell invasion assay using melanoma cells. I have found mixed information online about the thickness of the Matrigel matrix I should use, how to load it, and how cells should be passaged for different cancer cell types. Could you please advise or provide a resource? Thanks.
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Invasion assays for cancer cell analysis is an extensively studied and published application area where Matrigel matrix has been effectively used. To establish a reproducible assay many factors need optimization, some examples are:
- Cell seed
- Chemoattractant type and concentration
- Matrigel matrix protein concentration and volume (which relates to thickness)
- Duration of the assay
- Use (or not) of serum starvation prior to the assay
- Selection of the appropriate control
As a starting point we recommend coating an 8 micron 24-well insert (Corning Cat. No. 353097) with 0.1 mL of 200 to 300 μg/mL of Corning Matrigel matrix (Corning Cat. Nos. 354234 and 354230) per insert. Titration of the concentration and volume will help to tighten the conditions for your assay. There are many helpful documents on the Corning website, including:
- Corning Matrigel Matrix Frequently Asked Questions
- Considerations when Optimizing your Chemotaxis or Invasion Assay
Here are some examples from other literature:
- Current Opinion in Cell Biology, Vol 36, Oct 2015, 13-22. Modes of cancer cell invasion and the role of the microenvironment, G. Clark, D. Matic Vignjevic
- Cancer Res. 2016 Aug 15;76 (16):4595-7. Extracellular Matrix Invasion in Metastases and Angiogenesis: Commentary on the Matrigel “Chemoinvasion Assay”, Albini A.
Corning also has some optimized prepackaged solutions available for tumor cell invasion assays on both clear and Corning FluoroBlok™ light-blocking permeable supports. Here are a few great resources to get you started using the Matrigel matrix invasion assay products:
- Protocol: Cell Invasion Assay
- Corning® BioCoat™ Tumor Cell Invasion Systems, Frequently Asked Questions
There are also posters and webinars on this topic available on www.corning.com/lifesciences that you may find valuable.