If you’re a cell biologist, you’ll be aware of the direction of travel from complex, undefined, animal component cell culture reagents, to scaffolds free from animal components.

Basement membrane extracts, such as the eponymous Matrigel®, have supported decades of advancement in cell culture. Matrigel provides an environment that mimics in vivo conditions, is rich in ECM proteins such as laminin, collagen IV, and entactin, and supports pluripotency and differentiation whilst maintaining cell viability.

At Axol, we use Matrigel for much of our cell culture work; it has been cited in over 12000 publications and is suitable for use with a broad range of iPSC-derived cell types, making it a popular choice for labs with multiple applications.

The animal origin of reagents introduces potential variability and undefined components, which can be particularly problematic for translational or GMP-compliant workflows. Animal origin basement membrane replacements may vary in protein concentration and composition across different lots, making it difficult to standardize experiments and are usually not DNase or RNase-treated and so contain trace amounts of nucleic acids.

Looking into Chemically Defined and Synthetic Alternatives

As the field evolves, so do the substrate options. In recent years, a number of xeno-free and fully defined ECM alternatives that offer improved consistency and compatibility with automation have entered the market. These substrates are particularly useful when transitioning to clinical-grade workflows or when developing cell-based assays that require high reproducibility, such as for screening and drug discovery applications.

At Axol, we have been trialling a new material, Ceglu^TM a chemically defined scaffold for cell culture developed by SEKISUI CHEMICAL CO., LTD.

Ceglu^TM is a chemically synthesized, animal-free scaffold. The manufacturer’s claims state that the material offers excellent reproducibility and is ideal for culturing adherent cells such as iPSCs. Ceglu^TM is stable at room temperature and can be used in an easily scalable coating solution. Pre-coated multiwell plates are ready to use without any additional preparation requirements.

Early results of Ceglu^TM vs Matrigel trials

We have tested Ceglu^TM-coated plates for cardiomyocyte differentiation and with our neuronal stem cells for all stages of iPSC-derived cell production

The below image shows iPSC cells cultured on Corning® Matrigel® Growth Factor Reduced (GFR) Basement Membrane Matrix, Product Number 356231 and Ceglu^TM multiwell plate, Catalogue number ASPL060001.

Our initial results with Ceglu^TM look good, and in the coming months we will be trialling the substrate with more of our cell lines and workflows. Cell morphology is similar for cells cultured on Matrigel, with well-defined edges and no signs of spontaneous differentiation. Images were taken at 4x magnification.

We sometimes see cell peeling during long term cell culture, particularly when culturing neuronal cells. We are hopeful that the uniform topography of Ceglu^TM coated cell culture surfaces, will reduce issues with cell peeling.

Conclusion

There are several factors to consider when using an ECM for cell culture, such as cost, time, and ease of use. Chemically defined substrates such as Ceglu^TM are easy to store and handle, with fully defined components.

Ceglu^TM fits with our direction of travel towards xeno-free reagents and early results suggest that cell morphology is comparable with cells cultured on Matrigel. Over the coming months, we plan to trial Ceglu^TM with additional cell lines and compare its performance with other matrix materials.

Have you tried alternatives to Matrigel? We’d be interested to hear your thoughts.

Let’s build better models, together.