How to Handle Matrigel | Storage, Coating, and Thawing Tips | Corning

Corning Matrigel matrix has been trusted for over 35 years to support cutting-edge research in cell biology. The current list of 13,000+ citations for Matrigel matrix continues to grow, with researchers regularly finding new applications for this reconstituted extracellular matrix (ECM) from Engelbreth-Holm-Swarm (EHS) mouse sarcoma. The formulation has been used for metabolism and toxicology studies, as well as for the development of several types of tumor cell invasion assays and most recently as a staple in organoid models — in addition to countless other purposes.

There are a few strategies for getting the most out of Matrigel matrix. Here's a look at how to handle Matrigel matrix in its various applications, the best practices for doing so, and why it continues to be the most trusted and widely used extracellular matrix.

Selecting the Appropriate Matrigel Formulation

The standard Matrigel formulation (8-12 mg/mL) is suitable for the culture of polarized cells, such as epithelial cells. It promotes the differentiation of many cell types, including hepatocytes, neurons, beta-islets, mammary epithelial, endothelial, and smooth muscle cells. High concentration (HC) Matrigel (18-22 mg/mL) provides greater matrix stiffness and scaffold integrity. It's suitable in vivo cell delivery applications, such as plug assays in mice.

Use phenol red-free Matrigel matrix formulations for colorimetric or fluorescence assays and when estrogenic effects are a concern. Choose a growth factor reduced formulation for applications where a more highly defined basement membrane preparation is desired.

The hESC-qualified formulation of Matrigel matrix provides the reproducibility and consistency essential for human embryonic and induced pluripotent feeder-free stem cell culture.

Depending on your application and scale, you may want to consider cultureware precoated with Matrigel matrix. These include BioCoat dishes of 35-100 mm, BioCoat plates with six to 96 wells, and Matrigel matrix-3D plates with 96 or 384 wells to support spheroid and organoid models in a high throughput environment. 

How to Handle Matrigel matrix in Storage and Use

Store Matrigel matrix at -20°C in a non-frost-free freezer. To minimize temperature fluctuations, do not store it in a freezer door, a door that's opened frequently, or in a frost-free freezer.

Among the tips for handling Matrigel matrix, perhaps the most important is understanding that Matrigel matrix will start to form a gel at 10°C and will rapidly gel at temperatures greater than 22°C. Therefore, keep Matrigel matrix on ice at all times during handling.

Thaw Matrigel matrix vials overnight in ice at 2°C to 8°C. Use enough ice so the vial stays in ice for the entire thawing process, not in cold water or liquefied ice. Place the covered ice bucket in a cold room or toward the back of the refrigerator to minimize temperature changes. Pre-chill all pipette tips and labware that you plan to place into contact with Matrigel matrix.

Standard concentration Matrigel matrix is viscous but free-flowing at 4°C to 10°C, so use a positive displacement pipette or syringe for accurate measurement. This becomes especially critical for HC Matrigel matrix, which is very viscous and nontransparent. Standard concentration Matrigel matrix is transparent. Frozen or thawed vials of Matrigel matrix may range in color from straw yellow to dark red. This is due to carbon dioxide interacting with the bicarbonate buffer and phenol red (if present). These color variations are normal and will disappear upon equilibration with 5 percent CO2.

Once thawed, swirl in ice to promote an even distribution. Use the lot-specific protein concentration provided in the Certificate of Analysis to calculate the appropriate volume to use for specific applications. After the first thaw, make single-use aliquots of Matrigel matrix to avoid repeated freeze-thaw cycles. Use polypropylene or other freezer-compatible tubes and store at -70°C or -20°C. Be mindful of the lot-specific expiration date, which is two years from the date of manufacture.

Matrigel Matrix Handling Tips

Coating Surfaces with Matrigel matrix

Use greater than 3 mg/mL of Matrigel matrix to form a firm gel. For in vivo applications, do not dilute Matrigel matrix to a final concentration below 4 mg/mL. Thin layers (0.5 mm) of Matrigel matrix are generally recommended for cell attachment and proliferation applications — for example, with human pluripotent stem cells (hPSCs), neurons, and cardiomyocytes.

Coat surfaces with at least 50 uL/cm2 for thin gels. Cells may also be cultured inside thick layers (1 mm) of Matrigel matrix for 3D cell culture applications, such as ring assays and cell invasion studies. Coat surfaces with at least 150-200 uL/cm2 for thick gels. Extensive dilution (<3 mg/mL) will result in a thin, non-gelled protein layer that may be useful for cell attachment but may not be as effective in differentiation studies. See these FAQs for additional tips on working with Corning Matrigel matrix for specific applications, such as endothelial tube formation, preventing or inducing differentiation of ES/iPS cells, and organoid culture.

Dilute Matrigel matrix by adding it to an ice-cold solution of serum-free medium or PBS (not water). Mix gently by swirling or pipetting up and down. Then coat vessels on ice, on a Corning CoolRack or ThermalTray on ice, or on a Corning CoolBox. Protocols will vary depending on your application but typically involve incubating the vessel at 37°C for 30 minutes to allow for gel formation.

Use Matrigel-coated plates on the day of coating if possible. If not, you can store coated plates in an incubator at 37°C for up to one week in serum-free media. Alternatively, coated plates with a layer of serum-free medium can be sealed with parafilm and stored at 2°C to 8°C.

Evaluating Natural, Synthetic, or Mimetic Alternatives

While other biologically based ECM products may have similar properties, none can match Matrigel's 30 years of success across diverse cell culture applications. Corning Matrigel is the most widely used ECM because it can be trusted to work.

However, some applications may benefit from a more clearly defined matrix. Use the Corning Guide to Surface Selection by Cell Type to find the ECM proteins (natural, synthetic, or mimetic) that have been successfully used with specific cell types, complete with almost 400 references.

For additional tips and insights, download the Ultimate Guide to Matrigel Matrix or visit our Best Practices for Matrigel matrix YouTube Playlist on the Corning Life Sciences Channel.