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Please use this identifier to cite or link to this item: http://hdl.handle.net/123456789/14861

Title: Determination of Cytotoxic Activity of TNF .. a Using
Authors: Gamai E.Eld
Sameh E. EI-Shewemi
Issue Date: 2001
Abstract: Tumor n~crosis factor-alpha (TNF-a) is one of the major cytokines regulating a large spectrum of physiological and pathophysiological functions of the immune system. In this investigation, a colourimetric microassays and electron microscopy were used to study the cytotoxic effect of TNF-a on WEHI-13 VAR (mouse fibrosarcoma) as tumor target cells. Water-soluble tetrazolium (MTS/PMS) reduction as well as crystal violet (CV) staining assays were used to detect the cytotoxic activity induced by different TNFa concentrations (1, 10, 100, 1000, and 10,000 pg/ml) and incubation times (0, 1, 2, 4, 6, 9, and 20 hours). Cytotoxicity started to develop after 9 hrs of incubation and reached 100 % at 10,000 pg/ml after 20 hrs of incubation when detected by MTS/PMS assay and 86.1 % when detected by CV assay. Although, the MTS/PMS method expressed higher sensitivity than CV staining assay, the percentage of cytotoxicity induced by TNF-a was directly correlated to each other when detected by both colourimetric assays. The ultrastructural changes induced by TNF-a on target cells were mainly cytoplasmic rather than nuclear. These changes were also dependent on the dose and lor duration of exposure to TNF-a
URI: http://hdl.handle.net/123456789/14861
Appears in Collections:College of Applied Medical Sciences

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