Abstract：In recent years, high-efficiency, high-purity enrichment or capture of rare cells that containing less than 100 per milliliter of blood, and non-destructive release of captured target cells are hot research topics in the field of precise tumor treatment and early disease diagnosis. Among them, the enrichment of rare cells such as circulating tumor cells (CTCs) and fetal nucleated red blood cells (NRBCs) are the most extensive. Aiming at these two kinds of rare cells, we summarized the research progress in the field from the perspectives of separation and enrichment principle, experimental methods and separation efficiency, and analyzed the advantages and disadvantages of various methods. On the whole, the separation principle and methods of rare cells can be roughly divided into physical sorting method and immunoaffinity method. The former utilizes the physical characteristics of rare cells such as size and hydrodynamics. While in the later method, rare cells are isolated and enriched by utilizing the differences caused by the immunoaffinity reactions of specific receptors and antibodies, as well as the nucleic acid aptamers on the cell surfaces. In addition, we further introduced the physical properties of rare cells and the application of emerging technologies such as nanotechnology, microfluidics and single cell manipulation in the field of separation and enrichment of rare cells, and briefly compared various separation methods.
宋婉云, 王惠宇, 王明明, 张弢. 血液中稀有细胞分离和富集方法研究进展[J]. 中国生物医学工程学报, 2019, 38(4): 481-489.
Song Wanyun, Wang Huiyu, Wang Mingming, Zhang Tao. Progress of Separation and Enrichment of Rare Cells from Blood. Chinese Journal of Biomedical Engineering, 2019, 38(4): 481-489.
[1] Cohen SJ, Alpaugh RK, Gross S, et al. Isolation and characterization of circulating tumor cells in patients with metastatic colorectal cancer [J]. Clinical Colorectal Cancer, 2006, 6(2): 125-132. [2] Le Dieu R, Taussig D, Lister TA, et al. Negative immunomagnetic selection of T cells from peripheral blood of presentation AML specimens [J]. Journal of Immunological Methods, 2009, 348(1-2): 95-100. [3] Shizuru JA, Negrin RS, Weissman IL. Hematopoietic stem and progenitor cells: clinical and preclinical regeneration of the hematolymphoid system [J]. Annual Review of Medicine, 2005, 56: 509-538. [4] Plaks V, Koopman CD, Werb Z. Cancer. Circulating tumor cells [J]. Science, 2013, 341(6151): 1186-1188. [5] Balic M, Williams A, Lin H, et al. Circulating tumor cells: from bench to bedside [J]. Annual Review of Medicine, 2013, 64(1): 31-44. [6] Joosse SA, Gorges TM, Pantel K. Biology, detection, and clinical implications of circulating tumor cells [J]. EMBO Molecular Medicine, 2015, 7(1): 1-11. [7] Akolekar R, Beta J, Picclarelli G, et al. Procedure-related risk of miscarriage following amniocentesis and chorionic villus sampling: a systematic review and meta-analysis [J]. Ultrasound Obstet Gynecol, 2015, 45(1): 16-26. [8] Kolvraa S, Christensen B, Lykke-hansen L, et al. The fetal erythroblast is not the optimal target for non-invasive prenatal diagnosis: preliminary results [J]. The Journal of Histochemistry and Cytochemistry, 2005, 53(3): 331-336. [9] Arya SK, Lim B, Rahman AR. Enrichment, detection and clinical significance of circulating tumor cells [J]. Lab on a Chip, 2013, 13(11): 1995-2027. [10] Liu Shijie, Tian Zuhong, Zhang Lei, et al. Combined cell surface carbonic anhydrase 9 and CD147 antigens enable high-efficiency capture of circulating tumor cells in clear cell renal cell carcinoma patients [J]. Oncotarget, 2016, 7(37): 59877-59891. [11] Zhu Peixuan, Stanton ML, Castle EP, et al. Detection of tumor-associated cells in cryopreserved peripheral blood mononuclear cell samples for retrospective analysis [J]. Journal of Translational Medicine, 2016, 14(1): 198. [12] Riethdorf S, Fritsche H, Muller V, et al. Detection of circulating tumor cells in peripheral blood of patients with metastatic breast cancer: a validation study of the CellSearch system [J]. Clinical Cancer Research, 2007, 13(3): 920-928. [13] Wang Xu, Qian Ximei, Beitler JJ, et al. Detection of circulating tumor cells in human peripheral blood using surface-enhanced Raman scattering nanoparticles [J]. Cancer Research, 2011, 71(5): 1526-1532. [14] Baecher-Allan C, Wolf E, Hafler DA. Functional analysis of highly defined, FACS-isolated populations of human regulatory CD4+ CD25+ T cells [J]. Clinical Immunology, 2005, 115(1): 10-18. [15] Lagus TP, Edd JF. A review of the theory, methods and recent applications of high-throughput single-cell droplet microfluidics [J]. Journal of Physics D-Applied Physics, 2013, 46(11): 114005. [16] Dimmeler S, Burchfield J, Zeiher AM. Cell-based therapy of myocardial infarction [J]. Arteriosclerosis, Thrombosis, and Vascular Biology, 2008, 28(2): 208-216. [17] Chang L, Howdyshell M, Liao WC, et al. Magnetic tweezers-based 3D microchannel electroporation for high-throughput gene transfection in living cells [J]. Small, 2015, 11(15): 1818-1828. [18] Au S H, Storey BD, Moore JC, et al. Clusters of circulating tumor cells traverse capillary-sized vessels [J]. Proceedings of the National Academy of Sciences of the United States of America, 2016, 113(18): 4947-4952. [19] Hosokawa M, Hayata T, Fukuda Y, et al. Size-Selective Microcavity Array for Rapid and Efficient Detection of Circulating Tumor Cells [J]. Analytical Chemistry, 2010, 82(15): 6629-6635. [20] Wu CC, Hong LZ, Ou CT. Blood cell-free plasma separated from blood samples with a cascading weir-type microfilter using dead-end filtration [J]. Journal of Medical and Biological Engineering, 2012, 32(3): 163-168. [21] Li Songzhan, Gao Yifan, Chen Xiran, et al. Highly efficient isolation and release of circulating tumor cells based on size-dependent filtration and degradable ZnO nanorods substrate in a wedge-shaped microfluidic chip [J]. Biomedical Microdevices, 2017, 19(4): 93. [22] Yamada M, Nakashima M, Seki M. Pinched flow fractionation: continuous size separation of particles utilizing a laminar flow profile in a pinched microchannel [J]. Analytical Chemistry, 2004, 76(18): 5465-5471. [23] Lee D, Sukumar P, Mahyuddin A, et al. Separation of model mixtures of epsilon-globin positive fetal nucleated red blood cells and anucleate erythrocytes using a microfluidic device [J]. Journal of Chromatography A, 2010, 1217(11): 1862-1866. [24] Sollier E, Go DE, Che J, et al. Size-selective collection of circulating tumor cells using Vortex technology [J]. Lab on a Chip, 2014, 14(1): 63-77. [25] Warkiani ME, Guan Guofeng, Luan KB, et al. Slanted spiral microfluidics for the ultra-fast, label-free isolation of circulating tumor cells [J]. Lab on a Chip, 2014, 14(1): 128-137. [26] Mehendale N, Sharma O, D'costa C, et al. A Radial Pillar Device (RAPID) for continuous and high-throughput separation of multi-sized particles [J]. Biomedical Microdevices, 2017, 20: 6. [27] Park H, Kim D, Yun KS. Single-cell manipulation on microfluidic chip by dielectrophoretic actuation and impedance detection [J]. Sensors and Actuators B-Chemical, 2010, 150(1): 167-173. [28] Braschler T, Demierre N, Nascimento E, et al. Continuous separation of cells by balanced dielectrophoretic forces at multiple frequencies [J]. Lab On A Chip, 2008, 8(2): 280-286. [29] Moon HS, Kwon K, Kim SI, et al. Continuous separation of breast cancer cells from blood samples using multi-orifice flow fractionation (MOFF) and dielectrophoresis (DEP) [J]. Lab on a Chip, 2011, 11(6): 1118-1125. [30] Liu Lukan, Yang Kaiguang, Zhu Xudong, et al. Aptamer-immobilized open tubular capillary column to capture circulating tumor cells for proteome analysis [J]. Talanta, 2017, 175: 189-193. [31] Hou Shuang, Chen Jiefu, Song Min, et al. Imprinted nanovelcro microchips for isolation and characterization of circulating fetal trophoblasts: Toward noninvasive prenatal diagnostics [J]. ACS Nano, 2017, 11(8): 8167-8177. [32] Zhao Libo, Lu Yi-tsung, Li Fuqiang, et al. High-purity prostate circulating tumor cell isolation by a polymer nanofiber-embedded microchip for whole exome sequencing [J]. Advanced Materials, 2013, 25(21): 2897-2902. [33] Zhao Weian, Cui Cheryl H, Bose S, et al. Bioinspired multivalent DNA network for capture and release of cells [J]. Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(48): 19626-19631. [34] Dharmasiri U, Njoroge SK, Witek M A, et al. High-throughput selection, enumeration, electrokinetic manipulation, and molecular profiling of low-abundance circulating tumor cells using a microfluidic system [J]. Analytical Chemistry, 2011, 83(6): 2301-2309. [35] Shen Qinglin, Xu Li, Zhao Libo, et al. Specific capture and release of circulating tumor cells using aptamer-modified nanosubstrates [J]. Advanced Materials, 2013, 25(16): 2368-2373. [36] Liu Hongliang, Li Yingying, Sun Kang, et al. Dual-responsive surfaces modified with phenylboronic acid-containing polymer brush to reversibly capture and release cancer cells [J]. Journal of the American Chemical Society, 2013, 135(20): 7603-7609. [37] Deng Yuliang, Zhang Yu, Sun Shuai, et al. An integrated microfluidic chip system for single-cell secretion profiling of rare circulating tumor cells [J]. Scientific Reports, 2014, 4: 7499-7507. [38] Wang Huiyu, Yue Guofeng, Dong Chaoqun, et al. Carboxybetaine methacrylate-modified nylon surface for circulating tumor cell capture [J]. ACS Applied Materials & Interfaces, 2014, 6(6): 4550-4559. [39] Cao Jing, Zhao Xiaoping, Younis MR, et al. Ultrasensitive Capture, Detection, and Release of Circulating Tumor Cells Using a Nanochannel-Ion Channel Hybrid Coupled with Electrochemical Detection Technique [J]. Analytical Chemistry, 2017, 89(20): 10957-10964. [40] Nagy GR, Ban Z, Sipos F, et al. Isolation of epsilon-haemoglobin-chain positive fetal cells with micromanipulation for prenatal diagnosis [J]. Prenatal Diagnosis, 2005, 25(5): 398-402. [41] Earhart CM, Hughes CE, Gaster RS, et al. Isolation and mutational analysis of circulating tumor cells from lung cancer patients with magnetic sifters and biochips [J]. Lab on a Chip, 2014, 14(1): 78-88. [42] Byeon Y, Ki CS, Han KH. Isolation of nucleated red blood cells in maternal blood for Non-invasive prenatal diagnosis [J]. Biomedical Microdevices, 2015, 17: 118. [43] Ozkumur E, Shah AM, Ciciliano JC, et al. Inertial focusing for tumor antigen-dependent and -independent sorting of rare circulating tumor cells [J]. Science Translational Medicine, 2013, 5(179): 179ra47. [44] Wang Lixue, Asghar W, Demirci U, et al. Nanostructured substrates for isolation of circulating tumor cells [J]. Nano Today, 2013, 8(4): 347-387. [45] Chen Weiqiang, Weng Shinuo, Zhang Feng, et al. Nanoroughened surfaces for efficient capture of circulating tumor cells without using capture antibodies [J]. ACS Nano, 2013, 7(1): 566-575. [46] Wang Shutao, Liu Kan, Liu Jian, et al. Highly efficient capture of circulating tumor cells by using nanostructured silicon substrates with integrated chaotic micromixers [J]. Angewandte Chemie International Edition, 2011, 50(13): 3084-3088. [47] Ke Zunfu, Lin M, Chen JF, et al. Programming thermoresponsiveness of NanoVelcro substrates enables effective purification of circulating tumor cells in lung cancer patients [J]. ACS Nano, 2015, 9(1): 62-70. [48] Wang Lixue, Zhu Chuandong, Zheng Qin, et al. Preparation of homogeneous nanostructures in 5 minutes for cancer cells capture [J]. Journal of Nanomaterials, 2015, 2015: 1-6. [49] Xu Gangwei, Tan Yulong, Xu Tiegang, et al. Hyaluronic acid-functionalized electrospun PLGA nanofibers embedded in a microfluidic chip for cancer cell capture and culture [J]. Biomaterials Science, 2017, 5(4): 752-761. [50] Zhang Nangang, Deng Yuliang, Tai Qidong, et al. Electrospun TiO2 nanofiber-based cell capture assay for detecting circulating tumor cells from colorectal and gastric cancer patients [J]. Advanced Materials, 2012, 24(20): 2756-2760. [51] Huang Chao, Yang Gao, Ha Qing, et al. Multifunctional “smart” particles engineered from live immunocytes: toward capture and release of cancer cells [J]. Advanced Materials, 2015, 27(2): 310-313. [52] Nel I, Gauler TC, Bublitz K, et al. Circulating tumor cell composition in renal cell carcinoma [J]. PLoS ONE, 2016, 11(4): e0153018. [53] Zhang Tian, Boominathan R, Foulk B, et al. Development of a novel c-MET-based CTC detection platform [J]. Molecular Cancer Research: MCR, 2016, 14(6): 539-547. [54] Juan ML, Righini M, Quidant R. Plasmon nano-optical tweezers [J]. Nature Photonics, 2011, 5(6): 349-356. [55] Liao CJ, Hsieh CH, Wang HM, et al. Isolation of label-free and viable circulating tumour cells (CTCs) from blood samples of cancer patients through a two-step process: negative selection-type immunomagnetic beads and spheroid cell culture-based cell isolation [J]. RSC Advances, 2017, 7(47): 29339-29349. [56] Mazutis L, Gilbert J, Ung WL, et al. Single-cell analysis and sorting using droplet-based microfluidics [J]. Nature Protocols, 2013, 8(5): 870-891. [57] Ashkin A, Dziedzic JM, Bjorkholm JE, et al. Observation of a single-beam gradient force optical trap for dielectric particles [J]. Optical Letters, 1986, 11(5): 288-291. [58] Dufresne ER, Grier DG. Optical tweezer arrays and optical substrates created with diffractive optics [J]. Review of Scientific Instruments, 1998, 69(5): 1974-1977. [59] Garces-chavez V, Quidant R, Reece PJ, et al. Extended organization of colloidal microparticles by surface plasmon polariton excitation [J]. Physical Review B, 2006, 73(8): 085417. [60] Guck J, Ananthakrishnan R, Moon TJ, et al. Optical deformability of soft biological dielectrics [J]. Physical Review Letter, 2000, 84(23): 5451-5454. [61] Liberale C, Cojoc G, Bragheri F, et al. Integrated microfluidic device for single-cell trapping and spectroscopy [J]. Scientific Reports, 2013, 3: 1258. [62] Li Peng, Mao Zhang, Peng Zhangli, et al. Acoustic separation of circulating tumor cells [J]. Proceedings of the National Academy of Sciences of the United States of America, 2015, 112(16): 4970-4975. [63] Ding Xiaoyun, Peng Zhangli, Lin SC, et al. Cell separation using tilted-angle standing surface acoustic waves [J]. Proceedings of the National Academy of Sciences of the United States of America, 2014, 111(36): 12992-12997. [64] Parkinson DR, Dracopoli N, Petty BG, et al. Considerations in the development of circulating tumor cell technology for clinical use [J]. Journal of Translational Medicine, 2012, 10: 138.
