3D Cell Culture

What is 3d cell culture?

In drug development, cancer research, as well as stem cell studies, cell culture is an important and crucial technique. Currently, most cells are cultivated using 2 Dimensional (2D) procedures, but new and enhanced approaches that utilize 3D Cell Culture techniques provide strong proof that far more advanced experiments can be undertaken, providing valuable insights.

The cell environment can be modified through 3D cell culture procedures, to imitate the cell in vivo and to offer more precise data on cell-to-cell interactions, tumor properties, drug development, metabolic profiling, stem cell research, and other disease kinds.[1]

A 3D cell culture is an environment in which biological cells can grow or interact in all three dimensions with their surroundings. A 3D cell culture enables in vivo cells to develop in all directions, as would other 2D environments (e.g. Petri dish). Cell cultures are similar. These three-dimensional cultures normally grow in small capsules, bioreactors, or in 3D cell colonies, where cells can develop into spheroids.

Difference between 2d culture and 3d culture/ 2d cell culture vs 3d cell culture?

Since the early 1900s, 2D cell cultures have been used. In recent years, scientists have given a lot of attention to 3D cell culture techniques, as they might yield precise tissue models.

The basis of biological research is simple, cost-effective, and reproducible, classic or 2D cell culture. In the meantime, 3D cell culture systems are more complicated for more in vivo consistency. The fundamental adjustments in 3D cells – morphology, motility, and polarity all altered – are pretty obvious but can have significant secondary tissue-specific effects.

While 2D culture traditionally has been a reliable and successful method for the cultivation of basic cells, 3D cell culture today offers analysts in the laboratory new capabilities and results. Do you use this new biological research approach? Can 3D culture provide new analytical and production possibilities for your laboratory?

Morphology of cells –

In a 2D environment, body cells don’t proliferate, hence settings in 2D systems can’t imitate the conditions in vivo that cells encounter. Cells cultivated in 2D do not, therefore, offer a proper morphology of the cells.

Expression of genes and proteins

The gene and protein expression of the cells produced in 2D systems does not frequently reflect the patterns in vivo.

Culture exposure – 

All cells are equally exposed to the culture environment when cells are cultivated on a bottle or petri-dish and include all growth agents, chemicals or medications included in it. This is not the case with body cells — medicines and nutrients are not evenly distributed to all cells. The diffusion limitations normally observed in the body cannot be replicated by a 2D culture method; the external cells can exchange easy nutrients and waste products, while the center cells of the 3D cultivation can’t exchange them so effortlessly.

Drug Sensitivity – 

In a 2D system, drug sensitivity and treatment may be over-sensitive, either due to a lack of a diffusion gradient, or because certain genes are not expressed as in vivo. An exact medication response provision is not available in the 2D model.

Range of cells

2D cell cultures are also relatively inflexible, and cellular behaviors can be investigated only to a limited extent.

These are only some of the reasons why a 3D culture environment has gained momentum in recent years.

Is It Time to Start Transitioning From 2D to 3D Cell Culture?

In recent years, there has been great scientific and industrial interest in the development of in vitro-engineered three-dimensional (3D) tissue substitutes that more closely mimic human tissues for the testing of cosmetic and pharmaceutical products.

Methods for advancing research are provided by both 2D and 3D cell culture techniques. 3D cell culture, on the other hand, has demonstrated that technology has the ability to dramatically transform the way novel drug therapies are tested, diseases are modeled, stem cells are used, and organ transplants are performed. 

As 3D cell culture becomes more prevalent, the procedures will become more well-understood, allowing for the development of more complex technologies. Researchers that are currently using 2D cell culture models to evaluate new pharmacological therapies should seriously investigate 3D cell culture options.

The advantages of co-culturing cells in 3D outweigh those of 2D cell culturing, and as tissue engineering techniques advance, so will tumor models, cancer treatment approaches, and disease testing methodologies.

How to move from 2D culture to 3D culture

For decades, biologists have relied on traditional 2D-grown monolayers of cells, yet this is an artificial system that does not reflect physiological reality. More biologists have begun to use 3D cellular models in recent years, which offer cells with a microenvironment that is more analogous to their natural surroundings. These 3D model systems may now be created using a number of strategies, and they are still substantially accessible to the same perturbation procedures as cell monolayers.

3D cultures, on the other hand, provide a number of hurdles, including determining how to standardize their development and adapting fluorescent cell-based assays, as well as dealing with the large amounts of imaging data generated and establishing appropriate image analysis algorithms.

With so many options, you might be wondering if switching from 2D to 3D is suitable for you. Above all, you want to ensure that tissue conditions are precisely duplicated. Before you get started, there are a few things to think about. The scaffold should be designed in such a way that it facilitates the creation of an optimum ECM. It should also be able to regulate the flow of oxygen, carbon dioxide, nutrients, and trash. Microenvironmental elements should be chosen based on their impact on cellular dynamics. Changing the hardness of 2D culture dish plastic surfaces to the softness of ECM gels, for example, has a significant impact on cell adhesion, spreading, migration, and differentiation.

One issue with growing cells in 3D cultures is the absence of consistency, which makes data comparison difficult. This is why scientists are attempting to build standard and rapid protocols for drug screening, as well as repeatable 3D cell culture platforms.

The environment and study goal will determine whether you utilize a 2D or 3D culture. 2D research are sometimes more practicable, rapid, and economical, and can yield relevant results. In some circumstances, however, 3D research may be preferable since they produce more accurate results that can be more easily converted into clinical observations.


  • Jensen, C., & Teng, Y. (2020). Is It Time to Start Transitioning From 2D to 3D Cell Culture? Frontiers in Molecular Biosciences, 7. https://doi.org/10.3389/fmolb.2020.00033
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  • Sartori, S. Tissue engineering approaches in the design of healthy and pathological in vitro tissue models. Nat. Rev. Mol. Cell. Biol. 5, 1–22. https://doi.org/10.3389/fbioe.2017.00040 (2017).