Measurement of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction activity and lactate dehydrogenase release using MTT
Measurement of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction activity and lactate dehydrogenase release using MTT
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DOI：
10.1016/s0168-0102(00)00188-7
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发表时间：
2000-12-01
影响因子：
2.9
通讯作者：
Matsuki, N
中科院分区：
文献类型：
--
作者：
Abe, K;Matsuki, N
3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四氮唑（MTT）还原试验和乳酸脱氢酶（LDH）释放试验已被广泛用于评价培养中的细胞活力。MTT还原测定测量活细胞的氧化还原活性，而LDH测定测量从死细胞释放到培养基中的LDH的活性。本文介绍了一种快速、简便的方法，即用同一种染料MTT测定细胞MTT还原和LDH释放。用于测量LDH活性的底物混合物含有乳酸盐、B-烟酰胺腺嘌呤二核苷酸、1-甲氧基吩嗪硫酸甲酯、MTT和Triton X-100。当含有LDH的培养基与底物混合时，MTT转化成MTT甲瓒与LDH活性成比例。该方法已成功应用于评价叔丁基过氧化氢在培养的大鼠皮层星形胶质细胞和培养的大鼠海马神经元谷氨酸的毒性。该方法经济、方便，尤其适用于在同一培养液中测定细胞MTT还原和LDH释放。(C)2000 Elsevier Science爱尔兰有限公司和日本神经科学学会。All rights reserved.
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay and lactate dehydrogenase (LDH) release assay have been widely used for evaluating cell viability in culture. MTT reduction assay measures the redox activity of living cells, while LDH assay measures the activity of LDH released into the medium from dead cells. In this paper, we introduce a quick and simple method or measuring cellular MTT reduction and LDH release with the same dye, MTT. The substrate mixture for measuring LDH activity contained lactate, B-nicotinamide adenine dinucleotide, 1-methoxyphenazine methosulfate, MTT and Triton X-100. When the medium containing LDH was mixed with the substrates, MTT was converted into MTT formazan in proportion to LDH activity. This method was successfully applied for evaluating t-butyl hydroperoxide toxicity in cultured I at cortical astrocytes and glutamate toxicity in cultured rat hippocampal neurons. Our method is economical and convenient especially for measuring cellular MTT reduction and LDH release in the same culture. (C) 2000 Elsevier Science Ireland Ltd and the Japan Neuroscience Society. All rights reserved.