Abstract: Disclosed are a composition for NK (natural killer) cell cryopreservation and an NK cell cryopreservation formulation. This composition can exhibit cell surface marker expression and NK cell degranulation ability at at least the same level as formulations using commercially available cryopreservation media, even in low-concentration DMSO (dimethyl sulfoxide) not including HEPES (hydroxyethyl piperazine ethane sulfonic acid) or HES (hydroxyethyl starch), and can show a high cell yield and vastly superior ability of NK cells to kill cancer cells. Since a conventional cryoprotectant, showing side effects due to nonspecific toxicity, is not included or is comprised at a low concentration and is replaced with a nontoxic and biocompatible cryoprotectant, the composition can be administered to a patient immediately after thawing without additional culture or replacement of the solution with an injectable solution, and is thus usable for a pharmaceutical formulation.