Perfusion culture for live-cell imaging: A review

1. Perfusion vs static cell culture

This short review will present some technical aspects and challenges encountered in perfusion culture for live-cell imaging.

Live-cell imaging is a non destructive method which focuses on the observation of live cells and is widely used in cellular biology research and biomedical industry.

Nowadays, the most widely used cell culture system is the static culture, where the cells are cultivated inside Petri dishes or multiwall plates. In static culture, the culture medium is supplied in a batch-wised manner. Static cell culture systems have the advantage of being economical and easy to use.

But in term of long-term cell culture, they present important flaws. The first drawback is the high risk of contamination caused by the repeated manual interventions. The second one is the fluctuation of the cell environment due to the medium replacement process.

Compared to static cell culture, perfusion cell culture provides a more sterile environment and a more stable culture environment thanks to the continuous nutrient supply and waste removal. This allows more quantifiable cell environment.

Perfusion culture also allows long-term study under microscope. With live-cell imaging techniques, it is critical to be able to replace culture medium without opening the perfusion chamber in order.

WEBINAR | Dynamic cell culture, a review

For more insight into this topic, please refer to this webinar introducing the concept of dynamic cell culture by incorporating perfusion, and describing the benefits of fluid flow, mechanical tension and shear stress to in vitro cell models of physiology and disease. This webinar was performed by Lisa Muiznieks, research engineer held in May 2020.

Here is the free PDF version of the webinar.

Perfusion is an important tool for 3D cell culture. The goal of 3D cell culture is to obtain a microenvironment mimicking as closely as possible the in vivo microenvironment. Indeed, in their native environment, cells are submitted to various biological and physical cues, such as cell-cell interactions, signalling molecules and mechanics of the surrounding extra cellular matrix.

In 3D cell culture, cells are seeded on a 3D scaffold material. This scaffold makes difficult manual interventions such as medium changing. Perfusion systems can thus be useful to overcome this issue.

Perfusion cell culture is certainly more challenging than conventional static cell culture, but allows to have a more precise control over cell microenvironment, which makes it an interesting tool for a wide range of biological applications.

2. Perfusion chambers & microfluidic cell culture systems

Perfusion cell culture systems can be used both with traditional perfusion chambers or microfluidic chips.

Perfusion chambers are widely commercially available. They are generally specifically designed for live imaging. In order to perform perfusion cell culture, it is required to choose closed configuration chambers with fluid lines. Dedicated chamber holders can be found, allowing to use several chambers at the same time. For more information about perfusion chambers for imaging, see our dedicated short review.

Microfluidics is currently a promising and growing field for cell culture applications.

Specific physical phenomena occurring at micrometer scale, such as the creation of laminar flows and concentration gradients, can be used to precisely control the microenvironment of the scale.

Simple and robust microfluidic systems for cell culture under perfusion are commercially available, and a growing number of researchers are focusing on designing specific microfluidic chips for cell culture in 2D or 3D.

For more information about microfluidic perfusion cell culture, see our dedicated review and the bibliographic resources at the end of this review.

One critical aspects of a perfusion systems is the choice of the fluid delivery method. Several types of pumping mechanisms are available.

To go into methods and techniques of perfusion for live cell imaging in depth, see this short review.

Other reviews & tutorials about perfusion culture for live cell imaging:

• Application notes for live cell imaging
• General review about perfusion chambers
• Continuous perfusion 3D cell culture
• Microfluidic for cell biology: concepts & methodologies

Conclusion

In conclusion, advanced perfusion systems are revolutionizing live-cell imaging by allowing continuous nutrient and oxygen exchange, which is critical for maintaining cellular viability over prolonged observations. The implementation of laminar flow within these systems enables highly controlled fluid dynamics, minimizing cell disturbance and ensuring a stable environment that mimics physiological conditions. This precise flow is crucial for capturing subtle cellular responses and is invaluable for applications across cell biology, pharmacology, and regenerative medicine.

Elveflow stands out as a trusted scientific partner, offering high-performance microfluidic systems optimized for experimental versatility and precision. With extensive expertise in both laminar flow systems and microfluidics integration, Elveflow supports researchers in achieving reproducible and insightful results, facilitating breakthroughs across a wide array of cell-based studies. For scientists seeking to enhance their live-cell imaging capabilities, Elveflow provides not only state-of-the-art equipment but also a collaborative approach tailored to the specific needs of each research project.

References