Fabrication and Optimization of Edible, Biocompatible Gelatin hydrogels for Cultivated Meat Production

Abstract

Cultivated meat research has grown exponentially over the past decade due to increasing demand for more sustainable sources of protein. While various research groups and companies are positively contributing to the development of lab-grown meat production, this emerging field is experiencing several challenges related to cost, efficiency, environmental emissions and reliance on chemical cross-linkers in the fabrication process. Scaffolds are a promising tool in cultivated meat research to provide a rapid, cost-effective form of production. Hydrogels are extremely biocompatible due to their porosity, stiffness and their ability to hold media. In current research, gelatin has been used as a scaffolding material due to its versatility, low cost and biocompatibility; however, gelatin scaffold fabrication frequently involves cross-linking via cytotoxic chemical crosslinkers. Dehydrothermal treatment, an effective form of physical cross-linking, has been used to fabricate gelatin films that do not possess a porous structure. This thesis aims to address several challenges by investigating the potential of dehydrothermal treatment on porous gelatin hydrogels to form edible scaffolds suitable for bovine myoblast and adipocyte culture in vitro. In this study, hydrogel formation was accomplished without toxic chemical reagents, instead relying on safe dehydrothermal treatment for cross-linking of scaffolding material using an oven. Several assessments were conducted on the hydrogel scaffolds following the crosslinking procedure to determine suitability for cell culture. These assessments demonstrated that the hydrogels were stable in culture media for over fourteen days and supported high cell viability during this timeframe. Finally, in order to address challenges pertaining to co-culture, the fabricated hydrogels were seeded with bovine skeletal muscle cells and pre-adipocytes separately, then stacked in a layer-by-layer assembly. This stacking was tuned to meet different preferences, in ratios of 1:1 and 1:2. This technique demonstrates a method of making cultivated meat more accessible and reducing health effects from consumption.