Functionalization, preparation and use of cell-laden gelatin methacryloyl–based hydrogels as modular tissue culture platforms
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.
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Acknowledgements
The work presented by the authors was supported by the Australian Research Council (Future Fellowship awarded to T.J.K. and D.W.H.; Discovery Project grants awarded to D.L., T.J.K. and D.W.H.), the National Health and Medical Research Council of Australia (D.W.H.), Cancer Council Queensland (D.L.), the European Union (Marie Curie Fellowship PIOF-GA-2010-272286 to F.P.W.M.), the US National Science Foundation (EFRI-1240443 to A.K.), IMMODGEL (602694 to A.K.) and the US National Institutes of Health (EB012597, AR057837, DE021468, HL099073, AI105024 and AR063745 to A.K.). We also acknowledge the National Breast Cancer Foundation (IN-15-047 to D.W.H.) Foundation (IN-15-047 to D.W.H.) and an Australia-Harvard Fellowship (to A.K. and D.W.H.) from Harvard Club of Australia Foundation. Australia-Harvard Fellowship (to A.K. and D.W.H.) Harvard Club of Australia Foundation.
Author information
- Daniela Loessner and Christoph Meinert: These authors contributed equally to this work.
Authors and Affiliations
- Queensland University of Technology (QUT), Brisbane, Queensland, Australia Daniela Loessner, Christoph Meinert, Elke Kaemmerer, Laure C Martine, Peter A Levett, Travis J Klein, Ferry P W Melchels & Dietmar W Hutmacher
- Division of Biomedical Engineering, Department of Medicine, Biomaterials Innovation Research Center, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA Kan Yue & Ali Khademhosseini
- Division of Health Sciences and Technology, Harvard-Massachusetts Institute of Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA Kan Yue & Ali Khademhosseini
- Department of Orthopaedics, University Medical Center Utrecht, Utrecht, the Netherlands Ferry P W Melchels
- Institute of Biological Chemistry, Biophysics and Bioengineering, School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh, UK Ferry P W Melchels
- Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA Ali Khademhosseini
- Department of Bioindustrial Technologies, College of Animal Bioscience and Technology, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul, Republic of Korea Ali Khademhosseini
- Department of Physics, King Abdulaziz University, Jeddah, Saudi Arabia Ali Khademhosseini
- Australian Prostate Cancer Research Centre-Queensland, Translational Research Institute, Queensland University of Technology, Brisbane, Queensland, Australia Dietmar W Hutmacher
- George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA Dietmar W Hutmacher
- Institute for Advanced Study, Technische Universität München, Munich, Germany Dietmar W Hutmacher
- Daniela Loessner