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Interactions between tumor cells and immune cells are sparsely explored in 3D models, although occasional studies with 3D cell culture technologies have confirmed that tumor architecture influences cancer cell-immune system interactions. The development of technologies enabling controlled analysis of tumor-immune system interactions especially in 3D is highly challenging. Two photon polymerization (2PP) as a method for fabrication of various microstructures suitable for preparation of 3D tumor models finds applications in this context. In the present study 2PP technology was used to fabricate polymeric microfibers in 3D microspace that mimic fibers of extracellular matrix. UV-curable polymer OrmoComp (Micro Resist Technology GmbH) was irradiated by Newport Spirit ultrafast amplified laser operating at 520 nm. The fibers were made with one side anchored to the substrate with the supporting structure, whereas the other side was freely movable in space. The shape and curliness of fibers were adjusted by alternating parameters of fabrication, namely energy intensities and speed of fabrication. Before exposing fabricated fibers to live cells, microstructures were processed with KOH based treatment to enhance adhesion of cancer cells. Arrangement of the cancer cells in the network of polymeric fibers was visualized by confocal scanning microscopy. Cancer cells were able to proliferate and form spatial cellular clusters among polymeric fibers after several days of cultivation. At the same time they were available to immune cells that could be supplemented to the culture at any time. The results of the present study document feasibility to use 2PP technology to develop in vitro 3D models suitable for studies of tumor-immune cell interactions.
T. Teplicky,A. Mateasik,D. Kosnac,D. Chorvat, andB. Cunderlikova
"Fabrication of 3D fibre scaffolds for tumor-immune system interactions by two photon polymerisation", Proc. SPIE 11271, Laser 3D Manufacturing VII, 1127111 (2 March 2020); https://doi.org/10.1117/12.2545996
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T. Teplicky, A. Mateasik, D. Kosnac, D. Chorvat, B. Cunderlikova, "Fabrication of 3D fibre scaffolds for tumor-immune system interactions by two photon polymerisation," Proc. SPIE 11271, Laser 3D Manufacturing VII, 1127111 (2 March 2020); https://doi.org/10.1117/12.2545996