MEASUREMENT OF CYTO-TOXICITY BY TARGET-CELL RELEASE AND RETENTION OF THE FLUORESCENT DYE BIS-CARBOXYETHYL-CARBOXYFLUORESCEIN (BCECF)
MEASUREMENT OF CYTO-TOXICITY BY TARGET-CELL RELEASE AND RETENTION OF THE FLUORESCENT DYE BIS-CARBOXYETHYL-CARBOXYFLUORESCEIN (BCECF)
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DOI：
10.1016/0022-1759(88)90427-9
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发表时间：
1988-04-06
影响因子：
2.2
通讯作者：
HENKART, PA
中科院分区：
文献类型：
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作者：
KOLBER, MA;QUINONES, RR;HENKART, PA
为了利用新的可用的扫描显微荧光计的淋巴细胞介导的细胞毒性测定，一些市售的荧光染料，它们的适用性作为靶细胞标记物进行了筛选。其中之一，bis-carboxyethyl-carboxyfluorescein（BCECF），可用于约104个靶细胞的测定，并显示出比其他荧光素衍生物少得多的自发渗漏，同时仍然比51 Cr更快地渗漏。对于细胞毒性T淋巴细胞（CTL）的短期细胞毒性孵育（小于2小时），BCECF释放到上清液中的校正百分比与51 Cr平行。对于4小时测定，可以通过测量靶细胞保留的BCECF来定量细胞毒性。使用人CTL和自然杀伤（NK）细胞作为效应子，与各种淋巴细胞和成纤维细胞作为目标，在4小时的测定，BCECF保留技术被发现给细胞毒性值可比的51铬释放测定。与51 Cr释放试验相比，用扫描显微荧光计在微量滴定威尔斯孔中测量BCECF荧光的细胞毒性试验具有安全、经济和处理时间的优点。
In order to utilize a newly available scanning microfluorimeter for lymphocyte-mediated cytotoxicity assays, a number of commercially available fluorescent dyes were campared for their suitability as target cell markers. One of them, bis-carboxyethyl-carboxyfluorescein (BCECF), was useful for assays with about 104 target cells and showed substantially less spontaneous leakage than other fluorescenin derivatives, while still leaking more rapidly than 51Cr. For short cytotoxicity incubations (less than 2 h) with cytotoxic T lymphocytes (CTL), the corrected percentage BCECF release into the supernatant parallels that of 51Cr. For 4 h assays cytotoxicity could be quantitated by measuring the BCECF retained by target cells. Using human CTL and natural killer (NK) cells as effectors, with a variety of lymphoid cells and fibroblasts as targets in 4 h assays, the BCECF retention technique was found to give cytotoxicity values comparable to the 51Cr release assay. Cytotoxicity assays measuring BCECF fluorescence in microtiter wells with the scanning microfluorimeter offer advantages of safety, economy, and processing time compared with the 51Cr release assay.