Gelatin is a widely utilized bioprinting biomaterial due to its cell‐adhesive and enzymatically cleavable properties, which improve cell adhesion and growth. Gelatin is often covalently cross‐linked to stabilize bioprinted structures, yet the covalently cross‐linked matrix is unable to recapitulate the dynamic microenvironment of the natural extracellular matrix (ECM), thereby limiting the functions of bioprinted cells. To some extent, a double network bioink can provide a more ECM‐mimetic, bioprinted niche for cell growth. More recently, gelatin matrices are being designed using reversible cross‐linking methods that can emulate the dynamic mechanical properties of the ECM. This review analyzes the progress in developing gelatin bioink formulations for 3D cell culture, and critically analyzes the bioprinting and cross‐linking techniques, with a focus on strategies to optimize the functions of bioprinted cells. This review discusses new cross‐linking chemistries that recapitulate the viscoelastic, stress‐relaxing microenvironment of the ECM, and enable advanced cell functions, yet are less explored in engineering the gelatin bioink. Finally, this work presents the perspective on the areas of future research and argues that the next generation of gelatin bioinks should be designed by considering cell–matrix interactions, and bioprinted constructs should be validated against currently established 3D cell culture standards to achieve improved therapeutic outcomes.
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