Campbell John D M
Research and Development, Miltenyi Biotec GmbH, Friedrich Ebert Str. 68, 51429 Bergisch Gladbach, Germany.
Methods. 2003 Oct;31(2):150-9. doi: 10.1016/s1046-2023(03)00125-7.
The Cytokine Secretion Assay is an innovative method for analysing and enriching live cytokine-secreting cells. In this assay, a cytokine affinity matrix is built on the cell plasma membrane, which traps cytokines produced by the cell in response to specific stimuli. The specifically bound cytokine is then detected, and the cells optionally enriched, using fluorochrome-conjugated cytokine-specific antibodies and magnetic microbeads. This method allows extremely detailed phenotyping of live cells and the detection of cytokine responses at very low frequencies. Here, the latest cell staining and separation procedures are reviewed, with particular reference to the best application of the technology and troubleshooting in a variety of different situations.
细胞因子分泌测定法是一种用于分析和富集活的细胞因子分泌细胞的创新方法。在该测定法中，细胞因子亲和基质构建于细胞质膜上，它会捕获细胞响应特定刺激而产生的细胞因子。然后使用荧光染料偶联的细胞因子特异性抗体和磁性微珠检测特异性结合的细胞因子，并可选择富集细胞。该方法能够对活细胞进行极其详细的表型分析，并检测极低频率的细胞因子反应。本文综述了最新的细胞染色和分离程序，特别提及了该技术在各种不同情况下的最佳应用及故障排除方法。
