Accurate detection of carcinoma cells by use of a cell microarray chip.
Accurate detection of carcinoma cells by use of a cell microarray chip.
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DOI：
10.1371/journal.pone.0032370
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Kataoka M
中科院分区：
文献类型：
--
作者：
Yamamura S;Yatsushiro S;Yamaguchi Y;Abe K;Shinohara Y;Tamiya E;Baba Y;Kataoka M
循环肿瘤细胞的准确检测和分析在转移性癌症的诊断和治疗中起着重要的作用。细胞芯片用于检测白细胞中的加标癌细胞。该芯片由聚苯乙烯制成，具有20，944个微室（宽105 μm，深50 μm）;观察微室中白细胞单层的形成。使用培养的人T淋巴母细胞样白血病（CCRF-CEM）细胞来检查细胞微阵列芯片检测掺入癌细胞的潜力。将T淋巴母细胞样白血病悬浮液分散在芯片表面上，然后静置15分钟以使白细胞沉降到微室中。当总共约600，000个白细胞分散到细胞微阵列芯片上时，在每个微室中发现约29个白细胞。类似地，当使用从人全血中分离的白细胞时，当总共约1，800，000个白细胞被置于细胞微阵列芯片上时，约89个白细胞进入每个微室。清洗芯片表面后，将PE标记的抗细胞角蛋白单克隆抗体和APC标记的抗CD 326（EpCAM）单克隆抗体溶液分散在芯片表面上，并使其反应15分钟;然后使用微阵列扫描仪在20分钟内检测任何荧光阳性细胞。在使用掺入的癌细胞的实验中（NCI-H1650，0.01 - 0.0001%），用PE标记的抗细胞角蛋白单克隆抗体实现了癌细胞的准确检测。此外，通过用上述单克隆抗体进行双重染色来验证微室中的癌细胞。显示了细胞微阵列芯片用于检测CTC的潜在应用，从而证明了通过在单个癌细胞水平上对细胞角蛋白和EpCAM进行双重染色的准确检测。
Accurate detection and analysis of circulating tumor cells plays an important role in the diagnosis and treatment of metastatic cancer treatment. A cell microarray chip was used to detect spiked carcinoma cells among leukocytes. The chip, with 20,944 microchambers (105 µm width and 50 µm depth), was made from polystyrene; and the formation of monolayers of leukocytes in the microchambers was observed. Cultured human T lymphoblastoid leukemia (CCRF-CEM) cells were used to examine the potential of the cell microarray chip for the detection of spiked carcinoma cells. A T lymphoblastoid leukemia suspension was dispersed on the chip surface, followed by 15 min standing to allow the leukocytes to settle down into the microchambers. Approximately 29 leukocytes were found in each microchamber when about 600,000 leukocytes in total were dispersed onto a cell microarray chip. Similarly, when leukocytes isolated from human whole blood were used, approximately 89 leukocytes entered each microchamber when about 1,800,000 leukocytes in total were placed onto the cell microarray chip. After washing the chip surface, PE-labeled anti-cytokeratin monoclonal antibody and APC-labeled anti-CD326 (EpCAM) monoclonal antibody solution were dispersed onto the chip surface and allowed to react for 15 min; and then a microarray scanner was employed to detect any fluorescence-positive cells within 20 min. In the experiments using spiked carcinoma cells (NCI-H1650, 0.01 to 0.0001%), accurate detection of carcinoma cells was achieved with PE-labeled anti-cytokeratin monoclonal antibody. Furthermore, verification of carcinoma cells in the microchambers was performed by double staining with the above monoclonal antibodies. The potential application of the cell microarray chip for the detection of CTCs was shown, thus demonstrating accurate detection by double staining for cytokeratin and EpCAM at the single carcinoma cell level.
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DOI：
10.1186/bcr2131
发表时间：
2008
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Deng G;Herrler M;Burgess D;Manna E;Krag D;Burke JF
通讯作者：
Burke JF
影响因子：
3.8
作者：
Mostert, Bianca;Kraan, Jaco;Sleijfer, Stefan
通讯作者：
Sleijfer, Stefan
影响因子：
7.4
作者：
Yamamura, S;Kishi, H;Muraguchi, A
通讯作者：
Muraguchi, A
影响因子：
3.7
作者：
Yatsushiro S;Yamamura S;Yamaguchi Y;Shinohara Y;Tamiya E;Horii T;Baba Y;Kataoka M
通讯作者：
Kataoka M
影响因子：
10.3
作者：
Sieuwerts, Anieta M.;Kraan, Jaco;Bolt, Joan;van der Spoel, Petra;Elstrodt, Fons;Schutte, Mieke;Martens, John W. M.;Gratama, Jan-Willem;Sleijfer, Stefan;Foekens, John A.
通讯作者：
Foekens, John A.