Inaccuracies in MTS assays: major distorting effects of medium, serum albumin, and fatty acids
Inaccuracies in MTS assays: major distorting effects of medium, serum albumin, and fatty acids
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DOI：
10.2144/04373st05
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发表时间：
2004-09-01
期刊：
影响因子：
2.7
通讯作者：
Walker, AM
中科院分区：
文献类型：
--
作者：
Huang, KT;Chen, YH;Walker, AM
可溶性甲醛测定法广泛用于细胞数量评估。然而;在我们的手中，我们经常观察到实际细胞数量与测定读数不一致的情况。在这项研究中，我们已经确定(i)在常用培养基中获得的大部分读数可能是由培养基成分扩增引起的，这种方式无法通过仅培养基对照来纠正；（ii） 10%血清中存在的白蛋白可使测定吸光度降低50%，从而掩盖细胞数量的实际加倍；（三）后一种效应取决于脂肪酸的浓度。为了解决这些问题，我们开发了一种方案，可以提供完全代表细胞数量的一致读数，同时保留了可溶性甲醛测定法的一些原始优势。
Soluble formazan assays are widely used for cell number assessment. However; in our hands, we observed frequent occasions in which the actual cell number was at odds with the assay reading. In this study, we have determined that (i) a large proportion of the reading obtained in commonly used culture media can be caused by media component amplification of formazan production in a way that cannot be corrected for by media-only controls; (ii) the albumin present in 10% serum can reduce the assay absorbance by 50% so that an actual doubling of cell number can be obscured; and (iii) this latter effect is dependent on the concentration of fatty acids. To counter these problems, we have developed a protocol that gives consistent readings that are fully representative of cell number while retaining some of the original advantages of soluble formazan assays.