Method for Obtaining and Studying a Gel Agar-Based Medium to Preserve the Electrical Activity of Rat Brain Slices after their Long–Term Cryopreservation

Abstract

Cryopreservation biotechnology allows a long-term preservation makes it possible to preserve and subsequent recovery of biological objects for a long time. It This technology is used for creating is necessary to create a cryobanks. In this work, wWe have developeded a two-component freezing solution consisting of an artificial cerebrospinal fluid and agar in different concentrations. The effectiveness of the solution in terms offor long-term cryopreservation was investigated on surviving slices of the olfactory cortex of the brain of such non-hibernating animals – as rats. Changes Variations in the activities of AMPA and NMDA glutamatergic mechanisms in brain slices were studied as functional indicators of successful cryopreservation. The following agar Different concentrations of agar were used: 33%, 44% and 50%. At a concentration of 33% agar, AMPA hyperactivation and recovery of NMDA recovery mechanisms were observed. At a concentration of 44% agar, hyperactivation of both mechanisms occurred. A cComplete recovery of the activities of the AMPA and NMDA mechanisms after prolonged cryopreservation (–10°C, 52 days) was achieved at an agar concentration of 50%. The developed freezing agar-based freezing solution developed and studied by us does not contains no “heavy” protectors (DMSO), antibiotics, and cations, such as Ba2+ and Sr2+, which normally lead to an irreversible blockade of AMPA and NMDA mechanisms. Thus, an the agar-based freezingdeveloped solution solution helpscontributes to maintaining a high level of activity of AMPA and NMDA activity mechanisms in slices during their cryopreservation. The developed solution can be used to create a cryobank of nervous tissue.