The major limitation of 3D bioprinting is the availability of inks. In order to develop new ink formulations, both their rheological behavior to obtain the best printability and the target bio-printed objects conformities must be studied. In this paper, for the first time in our knowledge, the preparation and the characterization of novel ink formulations based on two natural biocompatible polysaccharides, chitosan (CH) and guar gum (GG), are presented. Five ink formulations containing different proportions of CH and GG were prepared and characterized in terms of rheological properties and solvent evaporation. Their printability was assessed (by varying the nozzle diameter, pressure and speed) using an extrusion-based 3D bioprinting process performed directly in air at 37 °C. Results showed that the incorporation of GG improved both the printability of the pure chitosan ink by increasing the viscosity of the solution and the shape fidelity by accelerating the solvent evaporation. We showed that the ink containing 15% (w/w) of GG and 85% (w/w) of CH had the best printability. This formulation was therefore used for the preparation of membranes that were characterized by infrared spectroscopy (FTIR) and X-Ray Diffraction (XRD) before and after gelation as well as for their mechanical properties (Young modulus, strength and strain at break). The optimal process printing parameters were determined to be: 27 G micronozzle, extrusion pressure below 2 bars and robot head speed between 20 and 25 mm/s. This novel ink formulation is a guideline for developing 2D scaffolds (such as auto-supported membranes) or 3D scaffolds for biomedical applications.