Hybrid double-spiral microfluidic chip for RBC-lysis-free enrichment of rare cells from whole blood
Hybrid double-spiral microfluidic chip for RBC-lysis-free enrichment of rare cells from whole blood
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用于全血中稀有细胞的无红细胞裂解富集的混合双螺旋微流控芯片
DOI：
10.1039/d2lc00713d
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发表时间：
2022
期刊：
影响因子：
6.1
通讯作者：
Yoshida Tomokazu
中科院分区：
文献类型：
--
作者：
Shirai Kentaro;Guan Guofeng;Meihui Tan;Xiaoling Peng;Oka Yuma;Takahashi Yusuke;Bhagat Ali Asgar S.;Yanagida Masatoshi;Iwanaga Shigeki;Matsubara Nobuaki;Mukohara Toru;Yoshida Tomokazu
通过使用循环肿瘤细胞（CTC）的微创液体活检进行药物选择和治疗监测有望在不久的将来实现。对于CTC的临床应用，从全血中简单、高通量、单步CTC分离而无需红细胞（RBC）裂解和离心仍然是一个关键的挑战。在这项研究中，我们开发了一种新的癌细胞分离芯片，“混合双螺旋芯片”，它涉及两个不同的迪恩流分级（DFF）分离模式的半和全迪恩循环的串联组合，这是一个混合的DFF分离模式超高通量血液处理在高精度和尺寸分辨率的分离。该芯片允许在30分钟内快速处理5 mL全血，而无需RBC裂解和离心。RBC和白色血细胞（WBC）的去除率分别超过99.9%和99%。加标A549癌细胞的平均回收率为87%，5 mL血液中低至200个细胞。该装置可以实现细胞数量的连续减少，从大约1010个全血细胞减少到108个细胞，随后减少到106个细胞的数量级。所开发的方法可以与使用成像流式细胞术测量所有回收的细胞相结合。作为概念证明，从转移性乳腺癌患者（N = 10，1-69个CTC/5 mL）和转移性前列腺癌患者（N = 10，1-39个CTC/5 mL）的血液中成功富集和计数CTC。我们相信，所开发的方法将有利于全血中罕见CTC的自动化临床分析。
Drug selection and treatment monitoring via minimally invasive liquid biopsy using circulating tumor cells (CTCs) are expected to be realized in the near future. For clinical applications of CTCs, simple, high-throughput, single-step CTC isolation from whole blood without red blood cell (RBC) lysis and centrifugation remains a crucial challenge. In this study, we developed a novel cancer cell separation chip, “hybrid double-spiral chip”, that involves the serial combination of two different Dean flow fractionation (DFF) separation modes of half and full Dean cycles, which is the hybrid DFF separation mode for ultra-high-throughput blood processing at high precision and size-resolution separation. The chip allows fast processing of 5 mL whole blood within 30 min without RBC lysis and centrifugation. RBC and white blood cell (WBC) depletion rates of over 99.9% and 99%, respectively, were achieved. The average recovery rate of spiked A549 cancer cells was 87% with as low as 200 cells in 5 mL blood. The device can achieve serial reduction in the number of cells from approximately 1010 cells of whole blood to 108 cells, and subsequently to an order of 106 cells. The developed method can be combined with measurements of all recovered cells using imaging flow cytometry. As proof of concept, CTCs were successfully enriched and enumerated from the blood of metastatic breast cancer patients (N = 10, 1–69 CTCs per 5 mL) and metastatic prostate cancer patients (N = 10, 1–39 CTCs per 5 mL). We believe that the developed method will be beneficial for automated clinical analysis of rare CTCs from whole blood.