Construction Of Sodium Alginate-gelatin@tea Polyphenol 3D Edible Scaffolds For Cell Culture Meat

Cell culture meat is a kind of meat product obtained by cultured animal muscle cells in vitro,which has shown great advantages in environmental protection,animal welfare and resource saving.Therefore,it is regarded as an efficient,green and sustainable new method of meat production.In the production process of cell culture meat,biological scaffolded is very important in order to construct structured cultured meat,which can not only provide cell support but also simulate extracellular matrix to promote cell proliferation and differentiation.Among them,porous scaffolders are conducive to cell expansion due to their internal interconnected network structure,and can also mimic the fibrous structure of skeletal muscle to prepare cultured meat pieces close to the size of real meat,which plays an important role in large-scale culture.In this study,a 3D edible scaffold with good biocompatibility was prepared by freeze-drying using sodium alginate and gelatin as the substrate,calcium chloride solution as the cross-linking agent and tea polyphenol as the coating material for the production of cell culture meat,and the details and results of the study are as follows:(1)Edible scaffolds prepared from sodium alginate,gelatin and tea polyphenol were successfully prepared.The surface and internal characteristics of the scaffolds were observed by scanning electron microscopy to investigate whether the pore structure of the scaffolds was suitable for cell culture;the physicochemical characteristics of the scaffolds were characterized using porosity,infrared,and swelling/degradation rates.It was observed that the scaffolds exhibited a porous laminar structure with tea polyphenol particles attached to the surface,which was favorable for cell growth.In addition,the prepared bio-scaffolds had good mechanical properties,and the IR results also demonstrated that tea polyphenols formed a coating on the scaffolds.(2)The S2-G1-TP0.1-3h bio-scaffold prepared by compounding sodium alginate and gelatin at a ratio of 2:1,tea polyphenol concentration of 0.1%and soaking time of 3 h were initially screened as the optimal conditions.The proliferation and differentiation characteristics of mouse myogenic cells on the scaffolds were characterized by live/dead cell staining and immunofluorescence staining;the formation of cellular myofibers on the scaffolds was observed by scanning electron microscopy,which was used to initially screen the ratio of scaffolds more suitable for cell culture.It was observed that the cells proliferated and differentiated well on the scaffold,and myofibers were formed on the scaffold.(3)The rabbit skeletal muscle myogenic cells were able to proliferate and differentiate on the scaffolds and eventually formed muscle tissue.The inoculation efficiency,metabolic viability,proliferation and differentiation ability,expression of proteins and genes related to myogenic differentiation,and microstructure of rabbit skeletal muscle myogenic cells on S2G1-TP0.1-3h scaffold were characterized.The results showed that the tea polyphenol coating facilitated the adhesion of rabbit skeletal muscle myogenic cells and the formation of cytoskeleton.There was a significant increase in both proteins and genes associated with myogenic differentiation of cells on the scaffold,and elongated fiber structures were also observed by scanning electron microscopy.(4)The rabbit-derived cell culture meat had similar appearance and textural characteristics to rabbit meat.The hardness,color,texture characteristics and water content of rabbit-derived cell-cultured meat were measured to investigate the similarities between cellcultured meat and natural meat products in terms of taste,texture characteristics and color.The results showed that the chromaticity values and textural characteristics such as hardness,chewiness,elasticity,cohesiveness and stickiness of the rabbit-derived cell-cultured meat were similar to those of rabbit meat.