Flow cytometric technique for quantitating cytotoxic response to photodynamic therapy
Flow cytometric technique for quantitating cytotoxic response to photodynamic therapy
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DOI：
10.1111/j.1751-1097.1996.tb03000.x
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发表时间：
1996-01-01
影响因子：
3.3
通讯作者：
Kennedy, JC
中科院分区：
文献类型：
--
作者：
Campbell, DL;Fisher, ME;Kennedy, JC
描述了一种简单的流式细胞术技术，用于快速测量细胞系统光敏化的多重细胞毒性反应。该技术对集落形成效率低的细胞系特别有用，需要进行非克隆性测定。该试验通过正向散射和侧向散射的门控将细胞大小的物体从细胞碎片中分离出来，通过钙黄蛋白AM阳性和乙二聚体-1阴性染色鉴定活细胞，并相对于聚苯乙烯珠的内标准测量细胞浓度，可以快速分析大量细胞。使用两种患者来源的小细胞肺癌细胞系NCI-H209和SV-E来测试该技术。这些细胞系对5-氨基乙酰丙酸诱导的原卟啉IX光敏反应的光敏存活曲线与光敏剂积累的程度相关。氚化胸腺嘧啶掺入试验与流式细胞术细胞毒性试验结果吻合较好。该技术可用于测量细胞系对光敏的细胞毒性反应，而不考虑其电镀效率。
A simple flow cytometric technique for rapid measurement of multilog cytotoxic responses to photosensitization of cellular systems is described. This technique is particularly useful for cell lines with a low colony-forming efficiency, for which a nonclonogenic assay is required. The assay separates cell-sized objects from cellular debris by gating on forward scatter versus side scatter, identifies viable cells by positive calcein AM and negative ethidium homodimer-1 staining and measures cell concentration relative to an internal standard of polystyrene beads, Large numbers of cells can be analyzed rapidly.Two patient-derived small cell lung cancer cell lines, NCI-H209 and SV-E, were used to test the technique. Photoradiation survival curves of the response of these cell lines to 5-aminolevulinic acid-induced protoporphyrin IX photosensitization correlated with the extent of photosensitizer accumulation. There was good agreement between the results obtained using the tritiated thymidine incorporation assay and the flow cytometric cytotoxicity assay. The technique can be used to measure cytotoxic responses to photosensitization of cell lines regardless of their plating efficiencies.