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~3H-TdR前标记法检测巨噬细胞细胞毒活性及其条件分析

A MODIFIED ASSAY FOR THE MACROPHAGE-MEDIATED CYTOTOXICITY BY USING THE ~3H-TdR LABELLED TARGET CELL AND ITS ASSESSMENT

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【作者】 董海东钱振超

【Author】 Dong Haidong & Qian Zhenchao Dept of pathophysiology, Dalian Medical College

【机构】 大连医学院病理生理学教研室大连医学院病理生理学教研室

【摘要】 本文介绍了以收集~3H-TdR前标记靶细胞为指标的检测巨噬细胞细胞毒活性的方法,并对其条件进行了分析。选用小鼠腹腔渗出性巨噬细胞,经24小时激活后,与~3H-TdR标记好的靶细胞共育24小时,收集细胞,测定细胞中残存放射量。从放射量(cpm)的减少来计算杀伤活性。实验表明,衣法结果准确、稳定、简便易行,而且无自发释放和~3H-TdR再利用问题。

【Abstract】 In this paper, we reported a modified method for measurement of the macrophage-mediated cytotoxicity. Thioglycollate-elicited routine peritoneal macrophages were activated by lymphokines and endotoxin in 24hr co-culture. The cells were collected on filter paper and detected by liquid scintilator. The cytotoxicity of macrophage could be represented by the deficit of cpm in filter paper (compared with target cell control). We found that there was a little possibility of reuse of the ~3H-TdR released from macrophage-killed labelled target cells in this assay. Our results showed tnat this method was good for macrophage-mediated cytotoxicity detection rather sensitive and reproducible, more convenient and less expensive.

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