title: Functionalization, preparation and use of cell-laden gelatin methacryloyl-based hydrogels as modular tissue culture platforms.
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.
authors.affiliation: Queensland University of Technology (QUT), Brisbane, Queensland, Australia.
authors.forename: Daniela
authors.initials: D
authors.lastname: Loessner
authors.affiliation: Queensland University of Technology (QUT), Brisbane, Queensland, Australia.
authors.forename: Christoph
authors.initials: C
authors.lastname: Meinert
authors.affiliation: Queensland University of Technology (QUT), Brisbane, Queensland, Australia.
authors.forename: Elke
authors.initials: E
authors.lastname: Kaemmerer
authors.affiliation: Queensland University of Technology (QUT), Brisbane, Queensland, Australia.
authors.forename: Laure C
authors.initials: LC
authors.lastname: Martine
authors.affiliation: Division of Biomedical Engineering, Department of Medicine, Biomaterials Innovation Research Center, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.|Division of Health Sciences and Technology, Harvard-Massachusetts Institute of Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
authors.forename: Kan
authors.initials: K
authors.lastname: Yue
authors.affiliation: Queensland University of Technology (QUT), Brisbane, Queensland, Australia.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Levett
authors.affiliation: Queensland University of Technology (QUT), Brisbane, Queensland, Australia.
authors.forename: Travis J
authors.initials: TJ
authors.lastname: Klein
authors.affiliation: Queensland University of Technology (QUT), Brisbane, Queensland, Australia.|Department of Orthopaedics, University Medical Center Utrecht, Utrecht, the Netherlands.|Institute of Biological Chemistry, Biophysics and Bioengineering, School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh, UK.
authors.forename: Ferry P W
authors.initials: FP
authors.lastname: Melchels
authors.affiliation: Division of Biomedical Engineering, Department of Medicine, Biomaterials Innovation Research Center, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.|Division of Health Sciences and Technology, Harvard-Massachusetts Institute of Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.|Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA.|Department of Bioindustrial Technologies, College of Animal Bioscience and Technology, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul, Republic of Korea.|Department of Physics, King Abdulaziz University, Jeddah, Saudi Arabia.
authors.forename: Ali
authors.initials: A
authors.lastname: Khademhosseini
authors.affiliation: Queensland University of Technology (QUT), Brisbane, Queensland, Australia.|Australian Prostate Cancer Research Centre-Queensland, Translational Research Institute, Queensland University of Technology, Brisbane, Queensland, Australia.|George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.|Institute for Advanced Study, Technische Universität München, Munich, Germany.
authors.forename: Dietmar W
authors.identifier: http://orcid.org/0000-0001-5678-2134
authors.initials: DW
authors.lastname: Hutmacher
mesh_terms: D000818:Animals; D001704:Biopolymers; D005780:Gelatin; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D008689:Methacrylates; D046509:Tissue Culture Techniques / Q000379:methods*; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D001704:Biopolymers; D020100:Hydrogels; D008689:Methacrylates; D005780:Gelatin; C031264:methacryloyl chloride