Hydrogels, typically favored for 3D printing due to their viscoelasticity, are now trending toward ecofriendly alternatives amid growing environmental concerns. In this study, we crafted cellulose-based hydrogels, specifically employing cellulose acetate sulfate (CAS). By keeping the acetyl group substitution degree (DSacetyl = 1.8) and CAS molecular weight constant, we varied rheological properties by adjusting sulfate group substitution (DSsulfate = 0.4, 0.7, and 1.0) and CAS concentration (2–5 wt %). Rheological characterizations, including shear-thinning, yield stress, and thixotropy, were performed to identify optimal conditions for formulating CAS hydrogel ink in direct ink writing for 3D printing under selected experimental conditions. Based on rheological findings, CAS hydrogels with DSsulfate 0.7 and concentration of 4 wt % was used for 3D printing, with subsequent evaluation of printing metrics. Additionally, the effect of ionic cross-linking using Ca2+ ions on the structural integrity of 3D-printed structures was evaluated, demonstrating effective preservation through reinforced polymer networks. The shrinking and swelling behaviors of the 3D-printed structures were also significantly affected by this ionic cross-linking. Building on these findings, this work could broaden the range of cellulose derivatives available for the preparation of cellulose-based hydrogels for 3D printing.