Functional recovery of adult brain tissue arrested in time during cryopreservation by vitrification

Cryopreserving the adult brain is challenging due to damage from ice formation, and traditional freezing methods fail to maintain neural architecture and function. Vitrification offers a promising alternative but has not been surveyed in the brain. Here, we demonstrate near-physiological recovery of the adult murine hippocampus after vitrification of brain slices and of the whole brain in situ. Key features of the hippocampus are preserved, including structural integrity, metabolic responsiveness, neuronal excitability, and synaptic transmission and plasticity. Notably, hippocampal long-term potentiation was well preserved, indicating that the cellular machinery of learning and memory remains operational. These findings extend known biophysical limits for cerebral hypothermic shutdown by demonstrating recovery after complete cessation of molecular mobility in the vitreous state. This suggests that the brain can be arrested in time and then reactivated, opening avenues for potential clinical applications. Significance StatementWhile the brain is considered exceptionally sensitive, we show that the hippocampus can resume normal electrophysiological activity after being rendered completely immobile in a cryogenic glass. The work extends known biophysical tolerance limits for the brain from the hypothermic to the cryogenic range and establishes a protocol for its long-term storage in a viable state.