Hydrophilic nanogels based on partially hydrolyzed poly(2-ethyl-2-oxazoline) were synthesized in dilute aqueous media in the presence of 1,6-hexanediol diglycidyl ether as a cross-linker. Nanogel formation was monitored by DLS and HSQC NMR spectroscopy, and the final nano-objects were characterized by DLS, TEM, AFM, and NanoSight analyses. Nanogels with a hydrodynamic radius of 78 nm exhibiting a slight positive surface charge were obtained. MTS assays (cell metabolic activity test) evidenced that nanogels were nontoxic in the investigated concentration range (i.e., 0.1 to 400 mu g/mL) and that no specific interaction with bovine serum albumin was observed.