Standardization of an in vitro drug screening assay by use of cryopreserved and characterized Pneumocystis carinii populations.
Standardization of an in vitro drug screening assay by use of cryopreserved and characterized Pneumocystis carinii populations.
复制标题
使用冷冻保存和特征化的卡氏肺囊虫群体进行体外药物筛选测定的标准化。
DOI：
10.1111/j.1550-7408.2001.tb00509.x
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Cushion,MT
中科院分区：
文献类型：
--
作者：
Collins,MS;Cushion,MT
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The feasibility of using cryopreserved organisms forin vitrodrug screening assays was evaluated to better standardized the ATP assay. ATP content and responses to standard anti-P. cariniicompounds were compared between freshly isolated and cryopreservedP. carinii. The ATP levels ofP. cariniiafter storage in liquid nitrogen (26 months) remained high throughout the evaluation period of 7 days. ATP pools from untreated organisms doubled by day 1 and maintained this level until day 6. Organism numbers, as determined by enumeration of nuclei, also doubled by 24 h, and slowly decreased after day 3. The ATP reduction in organisms treated with pentamidine was drastic. In both fresh and cryopreserved isolates ofP. carinii, peak metabolic activity occurred from days 1 to 3. Cryopreserved organisms initially had lower ATP levels than fresh, but by day 7, achieved similar results. Cryopreserved organisms were 50% more sensitive to pentamidine 2 µg/ml at day 1 with sensitivities closer to freshly isolated organisms at all other points. Counts of host cells, both live and dead, peaked at day 0 and declined throughout the 3-day assay period, while ATP levels increased to day 1 and remained stable thereafter. It is concluded that the use of cryopreservedP. cariniiallows characterization of the organism population prior to use in an entire series of experiments, which aids in standardization of the assay, as well as decreases the investigator's reliance on the availability of infected animals.