凝胶琼脂基大鼠脑切片长期冷冻保存电活动介质的制备与研究

A. Mokrushin
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引用次数: 0

摘要

低温保存技术允许长期保存,使生物物体的保存和后续恢复成为可能。如果这项技术用于建立一个冷冻银行是必要的。在这项工作中,我们开发了一种由不同浓度的人工脑脊液和琼脂组成的双组分冷冻溶液。我们在老鼠等非冬眠动物的大脑嗅皮质存活切片上研究了该溶液在长期低温保存方面的有效性。研究了AMPA和NMDA活性变化的脑切片谷氨酸能机制,作为低温保存成功的功能指标。使用不同浓度的琼脂:33%,44%和50%。琼脂浓度为33%时,观察到AMPA超激活和NMDA恢复机制的恢复。当琼脂浓度为44%时,这两种机制都发生了过度激活。在琼脂浓度为50%的情况下,经过长时间低温保存(-10°C, 52天),AMPA和NMDA机制的活性完全恢复。我们开发和研究的基于琼脂的冷冻液不含“重”保护剂(DMSO)、抗生素和阳离子,如Ba2+和Sr2+,这些通常会导致AMPA和NMDA机制的不可逆阻断。因此,琼脂冷冻液有助于在切片冷冻保存过程中保持AMPA和NMDA活性机制的高水平活性。开发的溶液可用于创建神经组织冷冻库。
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Method for Obtaining and Studying a Gel Agar-Based Medium to Preserve the Electrical Activity of Rat Brain Slices after their Long–Term Cryopreservation
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.
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