Label-free ferrohydrodynamic cell separation of circulating tumor cells.
Label-free ferrohydrodynamic cell separation of circulating tumor cells.
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DOI：
10.1039/c7lc00680b
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发表时间：
2017-09-12
期刊：
影响因子：
6.1
通讯作者：
Mao L
中科院分区：
文献类型：
--
作者：
Zhao W;Cheng R;Jenkins BD;Zhu T;Okonkwo NE;Jones CE;Davis MB;Kavuri SK;Hao Z;Schroeder C;Mao L
循环肿瘤细胞(CTCs)在癌症基础研究和临床应用中都具有重要意义。为了解决可用于基础和临床研究的有效CTCs的有限问题，有效地从血液中分离极其稀有的CTCs是至关重要的。铁流体动力细胞分离(FCS)是一种基于生物兼容铁液中细胞大小差异进行细胞分选的无标记方法，由于与处理临床样本相关的技术挑战，迄今无法从癌症患者的血液中浓缩低浓度的CTCs。在这项研究中，我们展示了一种层流微流控系统的开发，它能够以生物兼容的方式从患者的血液中富集稀有的CTCs，并且具有高通量(6mLh−1)和高回收率(92.9%)。通过验证的分析模型对FCS设备进行了系统优化，以确定能够处理临床上相关数量的血液的最佳磁场及其梯度、磁流体性质和细胞吞吐量。我们首先通过从未稀释的白细胞(WBC)中使用6个培养细胞系成功地分离低浓度(~100个细胞毫升−1)癌细胞的能力验证了该设备的能力，以每小时6毫升的吞吐量，平均癌细胞回收率为92.9%，分离的癌细胞平均纯度为11.7%。在~100个癌细胞−-1峰比下，癌细胞回收率分别为：H1299肺癌92.3±3.6%，A549肺癌88.3±5.5%，H3122肺癌93.7±5.5%，PC-3前列腺癌95.3±6.0%，乳腺癌94.7±4.0%，HCC1806乳腺癌93.0±5.3%。分离的癌细胞纯度分别为：H1299肺癌11.1%±1.2%，A549肺癌10.1±1.7%，H3122肺癌12.1±2.1%，PC-3前列腺癌12.8±1.6%，MCF-7乳腺癌11.9±1.8%，HCC1806乳腺癌12.2±1.6%。H1299细胞和HCC1806细胞的生物相容性研究表明，分离的癌细胞具有良好的短期生存能力，增殖正常，关键生物标记物的表达不受影响。然后，我们展示了从两名新诊断的晚期非小细胞肺癌(NSCLC)患者的血液样本中获得的CTCs的浓缩。虽然FCS仍处于开发的早期阶段，但由于其高回收率和良好的生物相容性，以及进一步优化和与其他分离方法整合的潜力，FCS可以成为四氯化碳分离的补充工具。
Circulating tumor cells (CTCs) have significant implications in both basic cancer research and clinical applications. To address the limited availability of viable CTCs for fundamental and clinical investigations, effective separation of extremely rare CTCs from blood is critical. Ferrohydrodynamic cell separation (FCS), a label-free method that conducted cell sorting based on cell size difference in biocompatible ferrofluids, has thus far not been able to enrich low-concentration CTCs from cancer patients’ blood because of technical challenges associated with processing clinical samples. In this study, we demonstrated the development of a laminar-flow microfluidic FCS device that was capable of enriching rare CTCs from patients’ blood in a biocompatible manner with a high throughput (6 mL h−1) and a high rate of recovery (92.9%). Systematic optimization of the FCS devices through a validated analytical model was performed to determine optimal magnetic field and its gradient, ferrofluid properties, and cell throughput that could process clinically relevant amount of blood. We first validated the capability of the FCS devices by successfully separating low-concentration (~100 cells mL−1) cancer cells using six cultured cell lines from undiluted white blood cells (WBCs), with an average 92.9% cancer cell recovery rate and an average 11.7% purity of separated cancer cells, at a throughput of 6 mL per hour. Specifically, at ~100 cancer cell mL−1 spike ratio, the recovery rates of cancer cells were 92.3 ± 3.6% (H1299 lung cancer), 88.3 ± 5.5% (A549 lung cancer), 93.7 ± 5.5% (H3122 lung cancer), 95.3 ± 6.0% (PC-3 prostate cancer), 94.7 ± 4.0% (MCF-7 breast cancer), and 93.0 ± 5.3% (HCC1806 breast cancer), and the corresponding purities of separated cancer cells were 11.1% ± 1.2% (H1299 lung cancer), 10.1 ± 1.7% (A549 lung cancer), 12.1 ± 2.1% (H3122 lung cancer), 12.8 ± 1.6% (PC-3 prostate cancer), 11.9 ± 1.8% (MCF-7 breast cancer), and 12.2 ± 1.6% (HCC1806 breast cancer). Biocompatibility study on H1299 cell line and HCC1806 cell line showed that separated cancer cells had excellent short-term viability, normal proliferation and unaffected key biomarker expressions. We then demonstrated the enrichment of CTCs in blood samples obtained from two patients with newly diagnosed advanced non-small cell lung cancer (NSCLC). While still at its early stage of development, FCS could become a complementary tool for CTC separation for its high recovery rate and excellent biocompatibility, as well as its potential for further optimization and integration with other separation methods.
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影响因子：
14.8
作者：
通讯作者：
--
影响因子：
6.1
作者：
Earhart CM;Hughes CE;Gaster RS;Ooi CC;Wilson RJ;Zhou LY;Humke EW;Xu L;Wong DJ;Willingham SB;Schwartz EJ;Weissman IL;Jeffrey SS;Neal JW;Rohatgi R;Wakelee HA;Wang SX
通讯作者：
Wang SX
影响因子：
4.6
作者：
Dhar M;Pao E;Renier C;Go DE;Che J;Montoya R;Conrad R;Matsumoto M;Heirich K;Triboulet M;Rao J;Jeffrey SS;Garon EB;Goldman J;Rao NP;Kulkarni R;Sollier-Christen E;Di Carlo D
通讯作者：
Di Carlo D
影响因子：
6.1
作者：
Huang, Song-Bin;Wu, Min-Hsien;Lee, Gwo-Bin
通讯作者：
Lee, Gwo-Bin
影响因子：
6.1
作者：
Kang, Joo H.;Krause, Silva;Ingber, Donald E.
通讯作者：
Ingber, Donald E.