Over the last few years, Deep Eutectic Solvents have gained popularity as a novel class of green solvents, due to their feasible synthesis and overall low production costs. The properties of glycerol (Gly)-based Deep Eutectic Solvents are frequently associated with the formation of an extended hydrogen bond network. In this study, two-dimensional Nuclear Magnetic Resonance (NMR) spectroscopy is employed to analyse the effect of glycerol oversaturation of the hydrogen bond acceptor, choline chloride (ChCl) on the structural arrangement of glyceline (molar ratio 1 : 2 ChCl:Gly), selected to represent Gly-based Deep Eutectic Solvents. The rearrangement of glycerol molecules, additionally trapping water molecules inside of isolated clusters, is revealed during a time-resolved analysis, performed in the presence of various fractions of water added to solvent. 200 % oversaturated Deep Eutectic Solvent (1 : 4 ChCl:Gly) is found to be a suitable cryoprotectant candidate, based on the revealed glycerol-water interactions.