Taramelli D, Holden H T, Varesio L
J Immunol Methods. 1980;37(3-4):225-32. doi: 10.1016/0022-1759(80)90309-9.
A short term microcytotoxicity assay is described for studying the activation of M0 by lymphokines. Proteose-peptone-induced macrophages were purified by adherence in flat-bottomed microtiter plates and then activated by MAF-containing supernatants for 18 h. 51Cr-labeled tumor target cells were added for an additional 18 h, and then the supernatants were harvested and the % isotope release quantitated. When endotoxin-free medium and FBS were used, we found that small amounts of LPS were absolutely required for macrophage activation in this assay. The advantages of this technique included (a) good reproducibility, (b) the requirement for small number of M0, and (c) the potential of standardizing the assay and thereby testing a large number of samples. Moreover, this assay may have particular value for investigations of M0 activation by exogenous stimuli, since M0 that were not pretreated with activating agents did not exhibit cytotoxicity.
本文描述了一种用于研究淋巴因子对M0细胞激活作用的短期微细胞毒性试验。用蛋白胨诱导的巨噬细胞通过贴壁于平底微量滴定板进行纯化，然后用含巨噬细胞激活因子（MAF）的上清液激活18小时。加入51Cr标记的肿瘤靶细胞，再培养18小时，然后收集上清液并定量测定同位素释放百分比。当使用无内毒素培养基和胎牛血清时，我们发现在该试验中巨噬细胞激活绝对需要少量的脂多糖（LPS）。该技术的优点包括：（a）良好的重复性；（b）所需的M0细胞数量少；（c）该试验有标准化的潜力，从而可以检测大量样本。此外，该试验对外源性刺激激活M0细胞的研究可能具有特殊价值，因为未经激活剂预处理的M0细胞不表现出细胞毒性。
