Calcein-Acetyoxymethyl Cytotoxicity Assay: Standardization of a Method Allowing Additional Analyses on Recovered Effector Cells and Supernatants

Simona Neri; Erminia Mariani; Alessandra Meneghetti; Luca Cattini; Andrea Facchini

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Calcein-Acetyoxymethyl Cytotoxicity Assay: Standardization of a Method Allowing Additional Analyses on Recovered Effector Cells and Supernatants

机译：钙黄绿素-乙酰氧基甲基细胞毒性测定：允许对回收的效应细胞和上清液进行额外分析的方法的标准化

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Cytotoxicity assays provide an in vitro evaluation of the lytic activity of NK and T cells against tumors or transformed cells. However, none of these methods allow the recovery of cells or supernatants after the assay. We standardized a microcytotoxicity test using calcein-acetoxymethyl (calcein-AM) dye that requires very small quantities of cells while maintaining the same sensitivity as the traditional 51Cr assay. The assay is applicable to resting as well as activated human effector cells and uses different targets such as human cell lines that are adherent or growing in suspension and resistant or sensitive. The most important feature of the method is the possibility of recovering cells and supernatants for additional analyses such as phenotyping and evaluation of soluble factors.

机译：细胞毒性测定提供了NK和T细胞针对肿瘤或转化细胞的裂解活性的体外评估。但是，这些方法均无法在测定后回收细胞或上清液。我们使用钙黄绿素-乙酰氧甲基（calcein-AM）染料对微细胞毒性测试进行了标准化，该染料需要非常少量的细胞，同时保持与传统 51 Cr测定相同的灵敏度。该测定法适用于静息的和激活的人类效应细胞，并使用不同的靶标，例如粘附或悬浮生长或耐药或敏感的人类细胞系。该方法的最重要特征是可以回收细胞和上清液以进行其他分析，例如表型分析和可溶性因子评估。

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《Clinical and diagnostic laboratory immunology》 |2001年第6期|共5页
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Simona Neri; Erminia Mariani; Alessandra Meneghetti; Luca Cattini; Andrea Facchini;
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3. Effects of concentrated supernatants recovered from Lactobacillus plantarum on Escherichia coli growth and on the viability of a human promyelocytic cell line [J] . Puertollano E., Puertollano M. A., Cruz-Chamorro L., Journal of applied microbiology . 2009,第4期

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5. Development of in vitro toxicity assays with a rainbow trout gill cell line and their use in determining the cytotoxicity and photocytotoxicity of polycyclic aromatic hydrocarbons individually and in a complex mixture [D] . Schirmer, Kristin 1997

机译：虹鳟鱼g细胞系体外毒性试验的发展及其在确定单个或复杂混合物中多环芳烃的细胞毒性和光细胞毒性中的用途
6. Calcein-Acetyoxymethyl Cytotoxicity Assay: Standardization of a Method Allowing Additional Analyses on Recovered Effector Cells and Supernatants [O] . Simona Neri, Erminia Mariani, Alessandra Meneghetti, 2001

机译：钙黄绿素-乙酰氧基甲基细胞毒性测定：允许对回收的效应细胞和上清液进行额外分析的方法的标准化
7. Calcein-Acetyoxymethyl Cytotoxicity Assay: Standardization of a Method Allowing Additional Analyses on Recovered Effector Cells and Supernatants [O] . Neri, Simona, Mariani, Erminia, Meneghetti, Alessandra, 2001

机译：钙黄绿素-乙酰氧基甲基细胞毒性测定：允许对回收的效应细胞和上清液进行额外分析的方法的标准化

Calcein-Acetyoxymethyl Cytotoxicity Assay: Standardization of a Method Allowing Additional Analyses on Recovered Effector Cells and Supernatants

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