Introduction to 3D Cell Culture
3D cell cultures are increasingly being used for basic research and drug discovery. Here you'll learn what 3D cell cultures are, why and how they are being used, and how to choose assays used with 3D models.
This guide provides an overview of 3D cultures and assays. Visit the 3D Assays page to view ordering information for a full list of assays optimized for use in 3D cultures.
What is 3D Cell Culture?
3D cell culture is a culture environment that allows cells to grow and interact with surrounding extracellular framework in three dimensions. This is in contrast with traditional 2D cell cultures in which cells are grown in a flat monolayer on a plate. 3D cell cultures can be grown with or without a supporting scaffold.
Scaffold 3D Cell Culture
3D cells may be cultured within a supporting scaffold to allow growth in all directions.
Popular types of scaffold include:
Hydrogels: Polymeric material containing a network of crosslinked polymer chains that can absorb and retain water. Hydrogels can be derived from animals (Matrigel®, collagen) or plants (alginate/agarose), or synthesized from chemicals (QGel® Matrix).
Inert matrices: Sponge-like membranes made of polystyrene which contain pores for cells to proliferate and grow.
Scaffold-Free 3D Cell Culture
3D cultures can be grown without the presence of a supporting scaffold. Scaffold-free methods rely on cells to self-assemble into clusters or spheroids.
Microfluidic Organ-on-a-Chip Models
An organ-on-a-chip is a 3D microfluidic cell culture chip engineered to mimic the physiology of an organ. 3D cells are grown in scaffolds within the chambers of a microchip. Tiny channels allow the flow of liquid (microliter to picoliter volumes) to transport and distribute nutrients or other chemicals throughout the cells.
Webinar: Overview of 3D Cell Culture Model Systems
Learn the pros and cons of commonly used 3D culture models and methods for detecting cell viability, apoptosis and oxidative stress in 3D cultures.
Why Use 3D Cell Culture?
For a long time, scientists have relied on flat 2D cell cultures grown on a plate to study cellular and disease mechanisms. 2D cell models are simple and cost-effective to culture and process. Within the last decade, however, 3D cell cultures have become increasing popular because they are more physiologically relevant and better represent in vivo tissue.
If you think about it, no cell types within our body grow as a monolayer independent of other cells or tissue. Instead, most cells naturally exist in complex 3D structures including different cell types within an extracellular matrix. The numerous cell-cell and cell-matrix interactions all have a profound effect on their behavior. In addition, 2D monolayers have uniform access to nutrients and oxygen, which is not the case in cell masses, such as tumors. 3D tumor spheroids are much more representative of in vivo tumors in which inner cells have less access to nutrients and oxygen compared to the outer layer, forming a natural gradient.
Here are some reasons to use 3D cell cultures:
- 3D cultures better mimic tissue-like structures
- Able to exhibit differentiated cellular function
- Possible to co-culture two or more different cell types
- Can simulate microenvironment conditions such as hypoxia and nutrient gradients
- Better predict in vivo responses to drug treatment
Feature Story: The Promises and Challenges of 3D Models
"That’s what we’re always doing with models: Trying to get them closer to being representative. I think everyone recognizes that 3D is a step forward."
In this feature article, you'll learn about the advantages and challenges of using 3D cell cultures.
Together, let's create a better world without animal testing. Let’s replace, reduce and refine.
3D Cell Culture Applications
3D cell models are increasingly being used to understand disease mechanisms and discover drug therapeutics. The process may involve deriving 3D cultures, such as cancer organoids, from patients. The 3D cultures can be used to screen for small molecule drugs or genetically manipulated to understand disease pathways. Compared to 2D cultures, 3D cell cultures more accurately predict the efficacy or toxicity of drug treatment.
Monitoring Biology in 3D Cultures
Data: 3D Cell Viability Assays
See how CellTiter-Glo® 3D Assay compares to other viability assays.
Article: Verifying Cell-Based Assays for Use with 3D Models
Assays originally intended for use with monolayer cell cultures need to be adapted to more complex 3D systems.
Story: Determining Cytotoxicity in 3D Liver Spheroids
Learn how the LDH-Glo™ Assay can help predict drug-induced liver injury in 3D microtissues.
Protocol: Assaying Cell Viability in Scaffold-Free 3D Culture
Learn how to measure cell viability from hanging drop culture using the CellTiter-Glo® 3D Assay and GloMax® Luminometer.
Webinar: Designing 3D Assays
Learn about factors to consider when designing and optimizing assays for 3D cultures.
Webinar: Method Validation and 3D Islet Models for Diabetes Research
Learn more about islet microtissues and their benefits for diabetes research.
Webinar: Liver Models and Lipid Metabolism: From Monolayer to Microtissue
Learn about 3D liver models and assays for measuring lipid metabolism.
Webinar: 3D Cell Culture and Assays Q&A
Our experts answer common questions on evaluating 3D assays for use with 3D cultures.
Case Study: 3D Assays for Ovarian Cancer Spheroids
Learn how a Yale researcher is using 3D cell viability assays to discover drugs and targeting methods for more efficient chemotherapy.