A COLORIMETRIC MICROTITER ASSAY FOR THE QUANTITATION OF CYTOKINE ACTIVITY ON ADHERENT CELLS IN TISSUE-CULTURE
A COLORIMETRIC MICROTITER ASSAY FOR THE QUANTITATION OF CYTOKINE ACTIVITY ON ADHERENT CELLS IN TISSUE-CULTURE
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DOI：
10.1016/0022-1759(92)90224-h
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发表时间：
1992-02-05
影响因子：
2.2
通讯作者：
KEISARI, Y
中科院分区：
文献类型：
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作者：
KEISARI, Y
开发了一种用于定量培养物中贴壁细胞的比色微量滴定法，其基于用市售染色试剂盒对细胞进行染色。 用Hemacolor试剂对粘附的L929、A375细胞系和人单核细胞进行染色，并用ELISA酶标仪在630 nm处通过比色法测定用0.5% SDS洗脱的颜色。 该方法能够检测细胞，并且线性高达3 × 10（4）L929细胞/孔。Hemacolor染色试验与结晶紫染色试验和MTT还原试验进行了比较，发现该方法灵敏、准确和可重复，并且具有在颜色洗脱之前能够对染色细胞进行显微镜检查的优点。 该方法适用于细胞毒性细胞因子的测定和粘附单核细胞的计数，也可用于细胞毒性药物的定量和病毒的细胞吞噬活性的测定。
A colorimetric microtiter assay was developed for the quantitation of adherent cells in culture, which is based on the staining of cells with a commercially available staining kit. Adherent L929, A375 cell lines and human monocytes were stained with Hemacolor reagents and the color eluted with SDS 0.5% was determined spectrophotometrically with an ELISA plate reader at 630 nm. The method enabled the detection of cells and was linear up to 3 x 10(4) L929 cells/well.The Hemacolor staining assay was compared to the crystal violet staining assay and the MTT reduction assay, and was found to be sensitive, accurate and reproducible, and has the advantage of enabling microscopic inspection of the stained cells prior to color elution. The assay was found to be suitable for the determination of cytotoxic cytokines, and the enumeration of adherent monocytes.This method might be also used for the quantitation of cytotoxic drugs, and the cytophatic activity of viruses.