Functionalization, preparation and use of cell-laden gelatin methacryloyl-based hydrogels as modular tissue culture platforms

Daniela Loessner, Christoph Meinert, Elke Kaemmerer, Laure C Martine, Kan Yue, Peter A. Levett, Travis J. Klein, Ferry P W Melchels, Ali Khademhosseini, Dietmar Werner Hutmacher

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Abstract

Progress in advancing a system-level understanding of the complexity of human tissue development and regeneration is hampered by a lack of biological model systems that recapitulate key aspects of these processes in a physiological context. Hence, growing demand by cell biologists for organ-specific extracellular mimics has led to the development of a plethora of 3D cell culture assays based on natural and synthetic matrices. We developed a physiological microenvironment of semisynthetic origin, called gelatin methacryloyl (GelMA)-based hydrogels, which combine the biocompatibility of natural matrices with the reproducibility, stability and modularity of synthetic biomaterials. We describe here a step-by-step protocol for the preparation of the GelMA polymer, which takes 1-2 weeks to complete, and which can be used to prepare hydrogel-based 3D cell culture models for cancer and stem cell research, as well as for tissue engineering applications. We also describe quality control and validation procedures, including how to assess the degree of GelMA functionalization and mechanical properties, to ensure reproducibility in experimental and animal studies.

Original languageEnglish
Pages (from-to)727-746
Number of pages20
JournalNature Protocols
Volume11
Issue number4
Early online date17 Mar 2016
DOIs
Publication statusPublished - Apr 2016

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    Loessner, D., Meinert, C., Kaemmerer, E., Martine, L. C., Yue, K., Levett, P. A., Klein, T. J., Melchels, F. P. W., Khademhosseini, A., & Hutmacher, D. W. (2016). Functionalization, preparation and use of cell-laden gelatin methacryloyl-based hydrogels as modular tissue culture platforms. Nature Protocols, 11(4), 727-746. https://doi.org/10.1038/nprot.2016.037