Preparation of CO₂-Triggered Extrudable Chitosan for Fat Production

Cultured ingredients are becoming increasingly important in the food industry. However, reproducing fat ingredients remains a major challenge. Hydrogel scaffolds are commonly used to culture cells for regenerative purposes. Among these, chitosan is a promising material, as it supports high cell viability. Still, its use is limited by conventional processing in acetic acid, whose residuals hamper cell availability and can modify food taste. Here, we propose a methodology for processing chitosan that enables CO₂-triggered structural modulation, supporting its application as a matrix for cultured food ingredients. For the first time, we add cells to the developed gel by standard mixing and using a newly developed method based on embedding predifferentiated adipocyte pellets. The findings reveal that the gel is stable and results in optimal cell viability when the cells are embedded in differentiated pellets. The system exhibits an effective release of lipids, thus validating its potential for application in edible fat-based ingredients. Moreover, the gel shows an interesting shear-thinning behavior, paving the road for extrusion-based processing.