Reversible growth arrest of 3D tumor spheroids stored in oxygen absorber-induced anoxia

Multicellular tumor spheroids models are of increasing interest in preclinical studies and pharmacological evaluation. However, their storage and transport is often a limitation because it requires adapted and expensive procedures. Here, we propose a very simple method to store 3D spheroids, using a procedure based on oxygen absorber-induced anoxia. We report that oxygen absorbers allow generating an anoxic environment for spheroid storage in culture plates. Oxygen absorber-induced anoxia fully and reversibly arrests spheroid growth for 4 days at 37°C and up to 18 days at 4°C. We then show that the response to etoposide is comparable in spheroids preserved in conditions of absorber-induced anoxia at 4°C and spheroids kept in normoxia at 37°C. These results represent a major improvement that should simplify the storage, transport and use of 3D spheroids.