Dynamic Microconfiner: Biomimetic 3D cell culture 

 novel Micronfiner for the first time allows to create a topological, mechanical and chemical controlled cell culture system for reproducible bio-mimetic culture conditions.

Challenge

In the past 60 years, we have observed a mind-blowing advancement in our quantitative understanding of life, ranging from bacteria over eukaryotic cells to tissue and even up to full organisms. This advancement has led to new medical treatments at a speed never seen before in human history. However, the impressive success masks a huge problem, pointed out in a series of articles published in Nature named “The replication crisis”. In the past years it has turned out that a major reason for this crisis is common practice of cultivating cells on 2D rigid substrates, which were found to lead to phenotypes showing largely different cellular behaviour in comparison to results found in the organism. in the past 20 years, it has been widely appreciated that controlling the culture conditions used in biological research is a key element to ensure reproducibility. Important aspects are dimensionality of the environment, stiffness of the substrate and as well as on the chemical functionalisation. Thus, the current state of the art biological research urgently requires controlled cultivation systems that allow for mimicking 3D situations, while ideally still compliant with 2D microscopy tools, and the possibility to control elastic properties and protein functionalization of the substrate.  

Our Solution

We have developed a cell confiner, which positions cells in a well-defined topological environment between two hydrogel (polyacrylamide) substrates (Figure 1). The instrument allows to dynamically adjust the distance between two substrates, such mimicking a classical 3D situation of migration through narrow gaps with the possibility to even create a moving and hence dynamically changing environment as found in the body. The distance between the substrates is controlled by a piezo motor, while a capacitive sensor ensures stability of the gap. The elastic properties of the top and bottom polyacrylamide gels that can be tuned between 100 Pa up to GPa, and the surface of each side can be chemically functionalized.

Figure 1: Left: Drawing of the prototype. Right: Example of LifeAct-GFP transfected neutrophils in a 3μm confinement. (upper) The cells feel both, a confinement from top and bottom, thus imitating a 3D confinement. (bottom) The PAA can be used to extract the traction forces applied by the cells. (Color code, 0-200Pa traction stress).

The System allows to provide constant and highly reproducible culture conditions for biomimetic 3D culture with controlled elastic and chemical surface properties.

Advantages

• Dynamic Microconfiner is compatible with high resolution microscopy
• High resolution microscopy is possible in a highly controlled biomimetic 3D cell culture
• Conditions like mechanical forces or chemical environment can be changed during imaging
• High precision hardware and an optimized design allows to mimic conditions in organisms
• A simple system mimicking the situation in an organism might highly enhance research output for the devlopment of new drugs

Applications

•  Research of cellular processes in a biomimetic 3D surrounding
• Pharma: improved evaluation of drug functionality
• Upscaling is planned in the lab, allowing higher sample throughput
• Drug screening

Development Status

Proof of concept with a working prototype. Cells were visualized under changing and well controlled conditions with high Resolution Microscopy. 

Patent Status

• Pending German Patent application

Contact

Dr. Martin Andresen
Patent Manager Life Sciences
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Tel.: +49 551 30724 150
Reference: BioC-2346-SUG

Tags: Microconfiner, biomimetic, reproducability, 3D cell culture, Light Sheet Imaging, Life science