Real-time impedance assay to follow the invasive activities of metastatic cells in culture
Real-time impedance assay to follow the invasive activities of metastatic cells in culture
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DOI：
10.2144/02334rr01
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发表时间：
2002-10-01
期刊：
影响因子：
2.7
通讯作者：
Giaever, I
中科院分区：
文献类型：
--
作者：
Keese, CR;Bhawe, K;Giaever, I
在此，我们展示了利用细胞-基质阻抗传感 (ECIS(TM)) 检测体外转移细胞侵袭活动的研究。该测定基于之前的显微镜观察，其中观察到添加到已建立的内皮细胞层上的转移细胞附着并侵入细胞层。人脐静脉内皮细胞（HUVEC）首先在小金电极上生长至汇合。添加 Dunning 鼠前列腺腺癌系列不同亚系（G、AT1、AT2、AT3、ML 和 MLL）的悬浮液后，跟踪这些电极的阻抗。对于高度转移的亚系，在受到攻击后一小时内，汇合的 HUVEC 层的阻抗大幅降低。弱转移亚系的影响不太明显，这种阻抗下降的程度和速率可能与测试的六个亚系中每一个的转移电位相关。实时测定在正常和低 (1%) 血清浓度下均有效，检测到的活性需要存在活的转化细胞。除了小鼠细胞系之外，在四种已建立的人类前列腺癌细胞系（DU145、PC3、TSU 和 PPC1）中也观察到了类似的行为。这些结果表明，这种基于 ECIS 的测定可用于原代人类培养物，以确定从活检中分离的细胞的转移能力。
Here we present research detecting the invasive activities of metastatic cells in vitro using electric cell-substrate impedance sensing (ECIS(TM)). The assay is based on previous microscopic observations, where metastatic cells added over established endothelial cell layers were observed to attach to and invade the cell layer Human umbilical vein endothelial cells (HUVECs) were first grown to confluence on small gold electrodes. The impedance of these electrodes was followed after the addition of suspensions of different sublines of the Dunning murine prostatic adenocarcinoma series (G, AT1, AT2, AT3, ML, and MLL). For highly metastatic sublines, within an hour after being challenged, the impedance of the confluent HUVEC layer was substantially reduced The effect of the weakly metastatic sublines was less pronounced, and the extent and the rate of this drop in impedance could be correlated with the metastatic potential of each of six sublines tested. The real-time assay is effective in both normal and low (1%) serum concentrations, and the detected activity requires the presence of viable transformed cells. In addition to the murine cell lines, similar behavior was observed using four established human prostatic cancer lines (DU145, PC3, TSU, and PPC1). These results suggest that this ECIS-based assay might be used with primary human cultures to establish the metastatic abilities of cells isolated from biopsies.