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与细胞及细胞库的遗传稳定性和保存相关的关键问题。

Key issues relating to the genetic stability and preservation of cells and cell banks.

作者信息

Simione F P

机构信息

American Type Culture Collection, Rockville, Maryland.

出版信息

J Parenter Sci Technol. 1992 Nov-Dec;46(6):226-32.

PMID:1474434
Abstract

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.

摘要

遗传稳定细胞的长期维持对于确保微生物学、细胞生物学和生物技术发展中的可重复性结果及连续性至关重要。作为活跃生长的培养物,细胞始终面临变化的风险,而对生物材料进行传代培养的必要性增加了遗传变化和污染的可能性。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兼容。保存活细胞最常用的方法是冷冻至低温以及冻干。主细胞库通常保存在液氮或类似温度下,而工作细胞库可以冷冻或冻干,并尽可能保存在更经济且易于管理的温度下。然而,低温技术可能会造成损害,导致遗传变化,或者在只有一小部分群体存活时产生潜在的选择。因此,一个良好的保存方案必须包括一个全面的细胞表征方案,该方案在保存细胞之前和之后都要应用,以确保在发生变化时能够检测到。长期稳定性的保证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尽量减少样本处理、验证和监测储存温度、提供备份库存以及对人员进行培训。细胞库还需要良好的编目和数据管理做法,以避免重复和错误识别,并确保对样本进行正确跟踪且便于获取。

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