Viable cultured neurons in ambient carbon dioxide and hibernation storage for a month

Neurobasal is a bicarbonate-buffered medium optimized for the growth of embryonic rat hippocampal neurons at pH 7.3 in 5% CO2. Neurons die within hours in this or in other 26 mM bicarbonate buffers when transferred to ambient CO2 (0.2%). Death is associated with a rapid rise in medium pH to 8.1. A new CO2-independent modification of Neurobasal (Hibernate E), when supplemented with B27, can maintain neuron viability for at least 2 days in ambient CO2. This same medium can also be used to store viable brain tissue for up to a month with refrigeration. These advances should facilitate studies of neuron physiology outside the incubator as well as storing and transporting neuronal tissue.