Isolation of circulating tumor cells using a microvortex-generating herringbone-chip
Isolation of circulating tumor cells using a microvortex-generating herringbone-chip
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DOI：
10.1073/pnas.1012539107
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发表时间：
2010-10-26
影响因子：
11.1
通讯作者：
Toner, Mehmet
中科院分区：
文献类型：
--
作者：
Stott, Shannon L.;Hsu, Chia-Hsien;Toner, Mehmet
存在于癌症患者血流中的罕见循环肿瘤细胞（CTC）为检测、表征和监测非血液学癌症提供了潜在的可获得来源。我们以前证明了微流体装置，CTC芯片，在捕获这些上皮细胞粘附分子（EpCAM）表达细胞使用抗体包被的微柱的有效性。在这里，我们描述了一种高通量的微流体混合装置，人字形芯片，或“HB芯片”，它提供了一个增强的CTC分离平台。HB-芯片设计通过产生微涡旋来应用血细胞的被动混合，以显著增加靶CTC与抗体包被的芯片表面之间的相互作用的数量。使用掺入对照血液中的限定数量的癌细胞验证了有效的细胞捕获，并且在前列腺癌患者的标本中证明了临床实用性。在15例转移性疾病患者中的14例（93%）中检测到CTC（中位数63 CTC/mL，平均值= 386 +/- 238 CTC/mL），并且在RNA分离和RT-PCR分析后容易鉴定肿瘤特异性TMPRSS 2-ERG易位。除了免疫荧光缀合的抗体之外，使用透明材料允许使用标准临床组织病理学染色对捕获的CTC进行成像。在患者样本的子集中，HB-芯片的低剪切设计揭示了CTC的微簇，这是以前未被认识到的肿瘤细胞聚集体，可能有助于癌症的血行播散。
Rare circulating tumor cells (CTCs) present in the bloodstream of patients with cancer provide a potentially accessible source for detection, characterization, and monitoring of nonhematological cancers. We previously demonstrated the effectiveness of a microfluidic device, the CTC-Chip, in capturing these epithelial cell adhesion molecule (EpCAM)-expressing cells using antibody-coated microposts. Here, we describe a high-throughput microfluidic mixing device, the herringbone-chip, or "HB-Chip," which provides an enhanced platform for CTC isolation. The HB-Chip design applies passive mixing of blood cells through the generation of microvortices to significantly increase the number of interactions between target CTCs and the antibody-coated chip surface. Efficient cell capture was validated using defined numbers of cancer cells spiked into control blood, and clinical utility was demonstrated in specimens from patients with prostate cancer. CTCs were detected in 14 of 15 (93%) patients with metastatic disease (median 63 CTCs/mL, mean = 386 +/- 238 CTCs/mL), and the tumor-specific TMPRSS2-ERG translocation was readily identified following RNA isolation and RT-PCR analysis. The use of transparent materials allowed for imaging of the captured CTCs using standard clinical histopathological stains, in addition to immunofluorescence-conjugated antibodies. In a subset of patient samples, the low shear design of the HB-Chip revealed microclusters of CTCs, previously unappreciated tumor cell aggregates that may contribute to the hematogenous dissemination of cancer.