Abstract:[Objective] This study aims to construct hydrogel scaffolds with both excellent mechanical properties and cell suitability for preparing high-quality structured cultured meat. [Method] Three biogenic protein gel materials (gelatin, collagen, and fibrinogen) and three polysaccharide gel materials (sodium alginate, chitosan, and gellan gum) were selected to construct single network (SN) hydrogels and double network (DN) hydrogels by cross-linking a single material or two materials of different categories. The physical properties (hardness, elasticity, and degradation rate) of the prepared hydrogels were characterized. Moreover, the biological properties of the prepared hydrogels were characterized in terms of cell adhesion, proliferation, and differentiation rates on the hydrogel surface. The scaffold function of the prepared hydrogels was then verified by three-dimensional culture of muscle stem cells. [Result] DN hydrogels demonstrated high hardness, high elasticity, and slow biodegradation, with the physical properties superior to those of SN hydrogels. In DN hydrogels, the addition of sodium alginate impeded the adhesion, proliferation, and differentiation of cells, and thus the biocompatibility of DN hydrogels with sodium alginate as the first network material was the lowest. DN hydrogels with chitosan as the first network material had high biocompatibility. In particular, the DN hydrogels with collagen and chitosan promoted the three-dimensional proliferation and differentiation of muscle stem cells, in which the cells showed the highest spreading and proliferation rates, as well as the highest expression of myosin heavy chain. [Conclusion] The material suitable for three-dimensional culture of porcine muscle stem cells are selected for the preparation of DN hydrogels based on high cell compatibility. The DN hydrogels with collagen and chitosan can be used as a scaffold material for preparing high-quality structured cultured meat.
