For more insight into methods and techniques to perform cell perfusion, 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.
Discover how to do microfluidic perfusion for dynamic cell culture.
Two different perfusion modes can be used: recirculating or non-recirculating. In recirculating mode, a given volume of culture medium is recirculating throughout the perfusion system. It is used in order to keep the molecule secreted by the cells in the culture medium. If cell-cell chemical communication is not essential to the experiment or if these chemical cues have to be ruled out, non-recirculating perfusion can be used, allowing to permanently remove secreted factors, waste products and thus wash cells.
Hydrodynamic shear stress can be a limiting factor for perfusion cell culture. In order to eliminate its effects, it is possible to lower the perfusion flow rate. Some have demonstrated that the minimum shear stress shown to affect differentiation or proliferation when applied continuously is about 0.5 N/m2. On the other hand, some experiments take advantage of the phenomena, by using high level of shear stress to investigate for example endothelial cell function in in-vivo conditions, i.e. with shear stress conditions or to assay cell adhesion.
Another way to reduce shear stress is to use microfluidic chips with dedicated geometries, such as microfluidic chips including a porous membrane acting as a barrier for the medium flow.
Temperature is a critical parameter of the cell environment for long-term studies on live cells. Several temperature control systems designed to be used while performing live-cell imaging are commercially available.
Air bubbles have a strong detrimental effect on cell culture. First, bubble trapped inside tubing or microfluidic channels can obstruct fluid flow. Then, bubbles are known to kill cells at their gas-liquid interface. A good way to eliminate bubbles is to apply a high flow rate inside the perfusion system prior to seeding the cells. A soft surfactant medium, like SDS is very effective to release air bubble from perfusion chamber.
A wide range of perfusion chambers are commercially available. Live cell imaging experiments can be performed with conventional perfusion chambers, microslides or microfluidic chips. Depending on your experiment, some chambers will be more suited to your needs. Don’t hesitate to read our review about perfusion chambers for imaging for more technical information about the different types of perfusion chambers, and to check our short overview of the different perfusion chambers commercially available.
Again, the choice of a perfusion system depends on the type of experiment. For simple live cell imaging experiments, pressure-driven flow controller, syringe or peristaltic pumps can be used. For experiments requiring more control over cells environment, injection at very low flow rates or switch between different media/reagents, pressure-driven are more adapted.
Feel free to contact our team of experts if you need help choosing the system that best fits the need of your experiment!
For more tutorial about microfluidics, please visit our other tutorials here: «Microfluidics tutorials».
The photos in this article come from the Elveflow® data bank, Wikipedia or elsewhere if precised. Article written by Emmanuelle Nadal and Timothée Houssin.
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