Challenges In Generating And Optimizing A 3D Cell Culture Model

Growing cells in a 3D culture is more preferable than a 2D culture on a plate or a support matrix because 3D culture allow the cells to grow with better cell-to-cell interaction and develop into a microtissue structure, which almost resembles the natural cellular environment.

Dr. Terry Riss, who is a Senior Product Specialist, Cell Health at Promega Corp, explained at a recently held webinar, Overview of 3D Cell Culture Model Systems & Validating Cell-based Assays for Use with 3D Cultures, that although 3D cell cultures are very beneficial to create an environment that is physiological, in which cell-based experiments can be performed very accurately, there are a lot of challenges that scientists are facing to design different experimental assays due to the complex and technical nature of the microtissue created through 3D cultures due to reagents failing to penetrate the tissue and giving the desire effect. Riss made several recommendations about how to design and optimize experimental strategies to get the desired results in a 3D system.  Riss discussed different aspects of the conventional methods of developing a 3D culture namely the scaffold based method and non-scaffold based method, their advantages and disadvantages and how to use them according to the need of the planned experiment.  According to Riss, as the scaffold-based methods use hydrogels as collagen, alginate and synthetic polymers, alginate is more beneficial than collagen because it is from a plant source and has fewer contaminants than the collagen, which is from an animal source. 

Although collagen creates a more extracellular matrix like environment for the cells to grow, but the chances of culture being contaminated are more.  On the other hand, synthetic polymers can be very advantageous, but these polymers need to of inert nature.  Riss also discussed the non-scaffold based methods like micropatterned plates and low-adhesion plates and how they are beneficial in controlling the size of the tissue, automation ability and reproducibility. Riss also put a lot of emphasis on methods of optimizing an assay originally designed for 2D culture to be used more proficiently and be more viable in a 3D microtissue and lot of date was presented on different assays of cytotoxicity, caspases and cell viability.