Dry Eye Disease (DED) is a multifactorial condition characterized by insufficient tear film stability and ocular discomfort. Conventional artificial tears offer limited efficacy due to short precorneal residence time. This study aimed to develop and optimize ion-sensitive in situ gelling formulations based on low-acyl Gellan Gum (GG) and Arabinogalactan (AG) to enhance retention and therapeutic efficacy in DED. Various buffer systems were screened to identify optimal gelation conditions upon interaction with artificial tear fluid (ATF). Formulations were characterized for pH, osmolality, wettability, thermal behavior, viscosity, and viscoelastic properties. A Design of Experiments (DoE) approach was employed to understand the influence of GG and AG concentrations on rheological behavior. The selected formulation, GG(0.1%)/AG(0.2%), demonstrated a significant viscosity increase upon ATF dilution, suitable viscoelastic properties, enhanced mucoadhesion compared to hyaluronic acid, improved ferning patterns, no cytotoxic effects, and stability over time. In vivo studies in rabbits confirmed prolonged precorneal retention of the fluorescently labeled formulation. These results suggest that the GG/AG-based hydrogel is a promising strategy for improving the performance of artificial tears in DED treatment.