We recently finished our Ask the Expert discussion, “Everything you ever wanted to ask about Corning® Matrigel® Matrix”. During this Ask the Expert session, we had a wide range of questions and discussed everything from storage and handling recommendations to Matrigel for 3D bioprinting. Specific topics included Matrigel best practices for: culturing a variety of cell types, imaging, analytical methods, cell invasion assays, harvesting, and 3D culture.

For the past 30 years, Corning Matrigel matrix has been used by researchers across the globe in essential applications through to cutting-edge, life-changing research. The number of citations for Matrigel matrix recently climbed over 10,000 citations and spans applications areas from cancer research to stem cells, and from organoid cultures to neuroscience. Corning recently published “The Ultimate Guide to Matrigel matrix” to share some of the best tips, tricks and expert advice and provided two experts to answer Cell Culture Dish reader questions.

This Ask the Expert session, was hosted by Katie Slater and Paula Flaherty, part of an extensive team of scientists that manufacture, test and develop products for applications that are used to modulate the in vitro behavior of cells via extracellular matrix proteins, cell culture surfaces, media and cultureware design. Paula is a member of the Corning Life Sciences leadership team as Technology Manager and of the Discovery Labware R&D team since 1984. She has extensive experience in developing cell based assays. Katie, a Senior Scientist since 2000, is a subject matter expert for the Corning extracellular matrix product line, focusing on the isolation, manufacture and testing of Corning Matrigel matrix and other clinically important extracellular matrix proteins.

Below is a sneak peek of the discussion, for a full transcript, please see – Ask the Expert – Everything you ever wanted to ask about Corning® Matrigel® Matrix

Question:

My question is about the future potential for Matrigel. Specifically, I’m interested in how Matrigel might be used as a bioink for 3D bioprinting. Have people been bioprinting with their cells in a Matrigel mixture? If so, what kinds of results have been seen (any publications I could read?) Besides bioprinting, are there any other new 3D cell culture techniques on the horizon?

Answer:

Corning® Matrigel® is poised to play an integral role in many 3D cell culture techniques, including bio-printing. Scientists have been using it to print many different tissues types. Listed below is a table that summarizes articles that have been published in this space recently that use Corning Matrigel in 3D bioprinting. Other 3D techniques that use Matrigel matrix are microfluidics and organ-on-a-chip for scaffold systems. For scaffold-free systems, Corning provides spheroid plates where the user can generate and analyze 3D spheres formed by one or more cell types. There is often a cross use of spheroid plates with Matrigel matrix if the customer is interested in generating self-assembled 3D structures.

Please see full answer for table of related literature – /ask_the_expert/question-future-potential-matrigel-specifically-im-interested-matrigel-might-used-bioink-3d-bioprinting-people-bioprinting-cells-ma/

Question:

Our lab is using Corning Matrigel matrix to co-culture cells in 3D on a microfluidic chip. I read that we should be using a thick layer of Matrigel matrix, but I’ve noticed that the amount of Matrigel matrix keeps going down each day and I need to add more. Do you have any recommendations so I can avoid having to add Matrigel matrix or should I be adding something else instead?

Answer:

Combining microfluidics and extracellular matrices (ECM) has shown to be a promising system to create more in vivo-like 3D environments. Some publications have shown different methods to craft such environments. For example:

• Bruzewicz, et al. (Lab Chip. 2008 May;8(5):663-71. doi: 10.1039/b719806j. Epub 2008 Mar 20) have shown that using a soft-lithographic molding gel such as Matrigel matrix or Collagen to encapsulate cells in a microfluidic channel and chambers yielded a permeable system where media could flow to feed the encapsulated cells.
• Jang, et al. (ACS Appl Mater Interfaces. 2015 Feb 4;7(4):2183-8. doi: 10.1021/am508292t. Epub 2015 Jan 21) have applied flow across the bulk gel during the gelation process to orient the ECM components with the direction of the flow, compared with randomly cross-linked Matrigel matrix.
• Tumour-on-a-chip: microfluidic models of tumour morphology, growth and microenvironment is a recently published review article by Tsai, et al. (J R Soc Interface. 2017 Jun; 14(131): 20170137).

Understanding the biophysical cues of the 3D environment such as topography, stiffness, viscosity and porosity have shown to be important to mimic the in vivo environment. Modulating and tuning the tensile strength of the Matrigel matrix gel in a 3D environment may be beneficial to provide softer or stiffer gels to suit application need. Empirical studies may show that a stiffer gel (higher protein concentration), may reduce dilution of the gel caused by the flow in the microfluidic chip.

As we keep learning about these techniques and methods, we recommend you reach out to our global scientific support team to help you find the right solution for your work.

Question:

What analytical methods are used to evaluate 3D cultures in Matrigel matrix?

Answer:

Cell viability, immunofluorescence analysis and advanced imaging technologies are frequently used to interrogate Matrigel matrix enabled 3D cultures.

Viability can be measured via the detection of DNA synthesis in proliferating cells, based on the incorporation of 5-ethynyl-2′-deoxyuridine (EdU). A protocol can be found below:

A chemical method for fast and sensitive detection of DNA synthesis in vivo

The labs of Mina Bissell at Lawrence Berkeley National Laboratory and Joan Brugge at Harvard Medical School have published extensively on 3D models using Matrigel matrix and have included a widely used immunofluorescence analysis preparation method. A few protocols can be found below:

ErbB2, but not ErbB1, reinitiates proliferation and induces luminal repopulation in epithelial acini

Three-dimensional culture models of normal and malignant breast epithelial cells

Many labs have studied 3D architecture utilizing advanced imaging technologies. In the publication by Jorgens, et al. many of these methods were employed and are covered in the materials and methods section of the paper. Spheroid/organoid size and morphology, as well as cryogenic techniques, volume electron microscopy, and super-resolution light microscopy have been used to study phenotypic and functional attributes. A protocol can be found below:

Deep nuclear invaginations are linked to cytoskeletal filaments – integrated bioimaging of epithelial cells in 3D culture

Finally, recovery of cells from 3D Matrigel matrix cultures to be used for cell number determination, RNA isolation, and qPCR analysis can be accomplished using Corning cell recovery solution. Using the solution at low temperature (on ice) and applying mechanical disruption such as pipetting or the use of an orbital shaker will help de-polymerize the Matrigel matrix. Cell-cell interactions can be disrupted through the use of chelators and/or proteolytic enzymes such as Trypsin or Dispase.