mtt法作为细胞增殖和细胞杀伤的快速检测:在人外周血淋巴细胞(PBL)中的应用。

Allergie und Immunologie Pub Date : 1991-01-01
H Weichert, I Blechschmidt, S Schröder, H Ambrosius
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引用次数: 0

摘要

研究了比色MTT法测定人外周血淋巴细胞(PBL)增殖和细胞死亡的可能性。在100,000-800,000细胞/孔范围内,发现光信号(570 nm处的OD信号)与细胞数呈线性相关。有必要将细胞与MTT孵育至少2小时。经不同PHA浓度刺激后,发现[3H]胸腺嘧啶掺入与MTT测定有很好的相关性。台盼蓝排除法和MTT法测定道霉素的细胞毒性,两者之间也有很好的相关性。可见MTT法是测定人PBL细胞增殖和细胞死亡的一种合适的方法。该方法易于操作,可在较短时间内检测大量探针,且不需要放射性。对于有色产物的测量,可以使用普通的ELISA读取器。
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The MTT-assay as a rapid test for cell proliferation and cell killing: application to human peripheral blood lymphocytes (PBL).

The possibility to use the colorimetric MTT assay for measuring proliferation and cell death of human peripheral blood lymphocytes (PBL) was studied. In a range from 100,000-800,000 cells/well a linear correlation between the optical signal (OD signal at 570 nm) and the cell number was found. It is necessary to incubate the cells with the MTT at least 2 hours. After stimulation by different PHA concentrations a very good correlation between [3H] thymidine incorporation and MTT assay was found. A comparison of daunomycin cytotoxicity, measurement by trypan blue exclusion and MTT assay, gave also a good correlation between both methods. It can be pronounced that the MTT assay is a suitable method to measure cell proliferation and cell death of human PBL. The assay is easy to handle, a large number of probes can be assayed in a relatively short time and no radioactivity is necessary. For the measurement of the colored product a common ELISA reader can be used.

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