与细胞和细胞库的遗传稳定性和保存有关的关键问题。

F P Simione
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引用次数: 0

摘要

在微生物学、细胞生物学和生物技术的发展中,长期维持遗传稳定的细胞对于确保可重复的结果和连续性是重要的。作为积极生长的培养物,细胞不断处于变化的风险中,传代培养活物质的必要性增加了遗传变化和污染的机会。有许多技术可用于稳定活细胞;所采用的方法必须与培养物的预期用途相容。保存活细胞最常用的方法是冷冻至低温和冷冻干燥。主库存通常保持在液氮或类似的温度下,而工作库存可以冷冻或冷冻干燥,并在可能的情况下保持在更经济和易于管理的温度下。然而,低温技术可能会造成损害,导致基因变化,或者当只有一小部分种群存活时,可能会出现选择。因此,一个好的保存程序必须包括在保存细胞之前和之后应用的全面的细胞表征方案,以确保在发生变化时检测到它们。确保长期稳定性需要精心设计的保管和安全措施,包括通过精心设计的库存系统,验证和监测储存温度,提供备用库存和人员培训来最大限度地减少标本处理。细胞库还需要良好的编目和数据管理实践,以避免重复和错误识别,并确保对标本的适当跟踪和易于获取。
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Key issues relating to the genetic stability and preservation of cells and cell banks.

The long term maintenance of genetically stable cells is important for ensuring reproducible results and continuity in the advance of microbiology, cell biology and biotechnology. As actively growing cultures, cells are constantly at risk of changing, and the necessity for subculturing living materials increases the chances for genetic change and contamination. Many techniques are available for stabilizing living cells; the method employed must be compatible with the intended use of the culture. The most commonly utilized means of preserving living cells are by freezing to cryogenic temperatures, and freeze-drying. Master stocks are usually maintained at liquid nitrogen or comparable temperatures, while working stocks can be frozen or freeze-dried, and maintained at more economical and easily managed temperatures where possible. However, low temperature techniques may cause damage that can result in genetic change, or potential selection when only a small portion of the population survives. Therefore, a good preservation program must include a comprehensive cell characterization regimen that is applied both before and after preserving the cells to ensure that changes are detected when they do occur. Assurance of long term stability necessitates well designed safekeeping and security measures that include minimizing specimen handling through well designed inventory systems, validation and monitoring of storage temperatures, provisions for backup inventory, and training of personnel. Cell banking also requires good cataloguing and data management practices to avoid duplication and misidentification, and to ensure proper tracking of specimens and ease of access.

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