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Mesuring O2 pressure in cell culture using 3D printed collagen gels.

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Gelatin and collagen gels are good candidates for the elaboration of cell culture media. Collagen, by its self-association properties, forms the framework of most tissues. In a biomimetic approach, collagen associated with gelatin seems to be an interesting candidate to reconstitute the dermis.

3D laser microprinting, a specialty of LIPHY, is a particularly promising fabrication route for structuring three-dimensional (3D) gels with specific architectures. In this study, the use of an instrument commercialized by Microlight3D allowing to spatially control the cross-linking of proteins at the focal point of the micro-laser to obtain gels with cubic cells or with more open structures such as stacks of hollow cubes.

For this process, it is necessary to add a photoinitiator to the starting medium, such as a ruthenium complex (a metal element that is part of the platinum family). This initiator, which allows the solidification of the medium at desired locations, also has interesting optical properties with respect to oxygen. Its photoluminescence (process by which it absorbs photons when excited by an external light source and then re-emits photons, thus light) increases when the quantity of oxygen in its environment decreases and decreases when the quantity of oxygen present in the vicinity increases. Thus, by following the luminescence intensity of ruthenium on the 3D structures fabricated in the laboratory and colonized by living cells, it was possible to highlight locally (at the cell size scales) the activity of these cells when they migrate on the gel structures or when they divide on these same 3D structures. Indeed in this case their oxygen consumption can be followed by the decrease of the fluorescence intensity of ruthenium present in the 3D structures.

Figure 2: Testing the ruthenium sensitivity in the gelatin-collagen matrix for 02 tension changes using normoxic A (19 % O2) – hypoxic B (1% O2)

Brevet européen (Euro/PCT) N° 17719651.6 du 29 mars 2017

View online : 3D two-photon polymerization of smart cell gelatin – collagen matrixes with incorporated ruthenium complexes for the monitoring of local oxygen tensions. Boudewijn van der Sanden, Laetitia Gredy, Didier Wion & Olivier Stephan. Acta Biomaterialia 130 2021.