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AUTHOR Clua-Ferré, Laura and de Chiara, Francesco and Rodríguez-Comas, Júlia and Comelles, Jordi and Martinez, Elena and Godeau, Amelie Luise and García-Alamán, Ainhoa and Gasa, Rosa and Ramón-Azcón, Javier
Title Collagen-Tannic Acid Spheroids for β-Cell Encapsulation Fabricated Using a 3D Bioprinter [Abstract]
Year 2022
Journal/Proceedings Advanced Materials Technologies
Abstract Type 1 Diabetes results from autoimmune response elicited against β-cell antigens. Nowadays, insulin injections remain the leading therapeutic option. However, injection treatment fails to emulate the highly dynamic insulin release that β-cells provide. 3D cell-laden microspheres have been proposed during the last years as a major platform for bioengineering insulin-secreting constructs for tissue graft implantation and a model for in vitro drug screening platforms. Current microsphere fabrication technologies have several drawbacks: the need for an oil phase containing surfactants, diameter inconsistency of the microspheres, and high time-consuming processes. These technologies have widely used alginate for its rapid gelation, high processability, and low cost. However, its low biocompatible properties do not provide effective cell attachment. This study proposes a high-throughput methodology using a 3D bioprinter that employs an ECM-like microenvironment for effective cell-laden microsphere production to overcome these limitations. Crosslinking the resulting microspheres with tannic acid prevents collagenase degradation and enhances spherical structural consistency while allowing the diffusion of nutrients and oxygen. The approach allows customization of microsphere diameter with extremely low variability. In conclusion, a novel bio-printing procedure is developed to fabricate large amounts of reproducible microspheres capable of secreting insulin in response to extracellular glucose stimuli.
AUTHOR Gretzinger, Sarah and Beckert, Nicole and Gleadall, Andrew and Lee-Thedieck, Cornelia and Hubbuch, Jürgen
Title 3D bioprinting – Flow cytometry as analytical strategy for 3D cell structures [Abstract]
Year 2018
Journal/Proceedings Bioprinting
The importance of 3D printing technologies increased significantly over the recent years. They are considered to have a huge impact in regenerative medicine and tissue engineering, since 3D bioprinting enables the production of cell-laden 3D scaffolds. Transition from academic research to pharmaceutical industry or clinical applications, however, is highly dependent on developing a robust and well-known process, while maintaining critical cell characteristics. Hence, a directed and systematic approach to 3D bioprinting process development is required, which also allows for the monitoring of these cell characteristics. This work presents the development of a flow cytometry-based analytical strategy as a tool for 3D bioprinting research. The development was based on a model process using a commercially available alginate-based bioink, the β-cell line INS-1E, and direct dispensing as 3D bioprinting method. We demonstrated that this set-up enabled viability and proliferation analysis. Additionally, use of an automated sampler facilitated high-throughput screenings. Finally, we showed that each process step, e.g. suspension of cells in bioink or 3D printing, cross-linking of the alginate scaffold after printing, has a crucial impact on INS-1E viability. This reflects the importance of process optimization in 3D bioprinting and the usefulness of the flow cytometry-based analytical strategy described here. The presented strategy has a great potential as a cell characterisation tool for 3D bioprinting and may contribute to a more directed process development.