An Optically Induced Dielectrophoresis (ODEP)-Based Microfluidic System for the Isolation of High-Purity CD45neg/EpCAMneg Cells from the Blood Samples of Cancer Patients—Demonstration and Initial Exploration of the Clinical Significance of These Cells
Circulating tumour cells (CTCs) in blood circulation play an important role in cancer metastasis. CTCs are generally defined as the cells in circulating blood expressing the surface antigen EpCAM (epithelial cell adhesion molecule). Nevertheless, CTCs with a highly metastatic nature might undergo an...
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Published in | Micromachines (Basel) Vol. 9; no. 11; p. 563 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
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Basel
MDPI AG
31.10.2018
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Abstract | Circulating tumour cells (CTCs) in blood circulation play an important role in cancer metastasis. CTCs are generally defined as the cells in circulating blood expressing the surface antigen EpCAM (epithelial cell adhesion molecule). Nevertheless, CTCs with a highly metastatic nature might undergo an epithelial-to-mesenchymal transition (EMT), after which their EpCAM expression is downregulated. In current CTC-related studies, however, these clinically important CTCs with high relevance to cancer metastasis could be missed due to the use of the conventional CTC isolation methodologies. To precisely explore the clinical significance of these cells (i.e., CD45neg/EpCAMneg cells), the high-purity isolation of these cells from blood samples is required. To achieve this isolation, the integration of fluorescence microscopic imaging and optically induced dielectrophoresis (ODEP)-based cell manipulation in a microfluidic system was proposed. In this study, an ODEP microfluidic system was developed. The optimal ODEP operating conditions and the performance of live CD45neg/EpCAMneg cell isolation were evaluated. The results demonstrated that the proposed system was capable of isolating live CD45neg/EpCAMneg cells with a purity as high as 100%, which is greater than the purity attainable using the existing techniques for similar tasks. As a demonstration case, the cancer-related gene expression of CD45neg/EpCAMneg cells isolated from the blood samples of healthy donors and cancer patients was successfully compared. The initial results indicate that the CD45neg/EpCAMneg nucleated cell population in the blood samples of cancer patients might contain cancer-related cells, particularly EMT-transformed CTCs, as suggested by the high detection rate of vimentin gene expression. Overall, this study presents an ODEP microfluidic system capable of simply and effectively isolating a specific, rare cell species from a cell mixture. |
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AbstractList | Circulating tumour cells (CTCs) in blood circulation play an important role in cancer metastasis. CTCs are generally defined as the cells in circulating blood expressing the surface antigen EpCAM (epithelial cell adhesion molecule). Nevertheless, CTCs with a highly metastatic nature might undergo an epithelial-to-mesenchymal transition (EMT), after which their EpCAM expression is downregulated. In current CTC-related studies, however, these clinically important CTCs with high relevance to cancer metastasis could be missed due to the use of the conventional CTC isolation methodologies. To precisely explore the clinical significance of these cells (i.e., CD45neg/EpCAMneg cells), the high-purity isolation of these cells from blood samples is required. To achieve this isolation, the integration of fluorescence microscopic imaging and optically induced dielectrophoresis (ODEP)-based cell manipulation in a microfluidic system was proposed. In this study, an ODEP microfluidic system was developed. The optimal ODEP operating conditions and the performance of live CD45neg/EpCAMneg cell isolation were evaluated. The results demonstrated that the proposed system was capable of isolating live CD45neg/EpCAMneg cells with a purity as high as 100%, which is greater than the purity attainable using the existing techniques for similar tasks. As a demonstration case, the cancer-related gene expression of CD45neg/EpCAMneg cells isolated from the blood samples of healthy donors and cancer patients was successfully compared. The initial results indicate that the CD45neg/EpCAMneg nucleated cell population in the blood samples of cancer patients might contain cancer-related cells, particularly EMT-transformed CTCs, as suggested by the high detection rate of vimentin gene expression. Overall, this study presents an ODEP microfluidic system capable of simply and effectively isolating a specific, rare cell species from a cell mixture. Circulating tumour cells (CTCs) in blood circulation play an important role in cancer metastasis. CTCs are generally defined as the cells in circulating blood expressing the surface antigen EpCAM (epithelial cell adhesion molecule). Nevertheless, CTCs with a highly metastatic nature might undergo an epithelial-to-mesenchymal transition (EMT), after which their EpCAM expression is downregulated. In current CTC-related studies, however, these clinically important CTCs with high relevance to cancer metastasis could be missed due to the use of the conventional CTC isolation methodologies. To precisely explore the clinical significance of these cells (i.e., CD45 neg /EpCAM neg cells), the high-purity isolation of these cells from blood samples is required. To achieve this isolation, the integration of fluorescence microscopic imaging and optically induced dielectrophoresis (ODEP)-based cell manipulation in a microfluidic system was proposed. In this study, an ODEP microfluidic system was developed. The optimal ODEP operating conditions and the performance of live CD45 neg /EpCAM neg cell isolation were evaluated. The results demonstrated that the proposed system was capable of isolating live CD45 neg /EpCAM neg cells with a purity as high as 100%, which is greater than the purity attainable using the existing techniques for similar tasks. As a demonstration case, the cancer-related gene expression of CD45 neg /EpCAM neg cells isolated from the blood samples of healthy donors and cancer patients was successfully compared. The initial results indicate that the CD45 neg /EpCAM neg nucleated cell population in the blood samples of cancer patients might contain cancer-related cells, particularly EMT-transformed CTCs, as suggested by the high detection rate of vimentin gene expression. Overall, this study presents an ODEP microfluidic system capable of simply and effectively isolating a specific, rare cell species from a cell mixture. |
Author | Wang, Hung-Ming Hsieh, Chia-Hsun Chiu, Tzu-Keng Wu, Min-Hsien Hung, Feng-Chun Liao, Chia-Jung Chou, Wen-Pin Zhu, Yu-Xian |
AuthorAffiliation | 2 Division of Haematology/Oncology, Department of Internal Medicine, Chang Gung Memorial Hospital (Linko), Taoyuan City 33302, Taiwan; wisdom5000@gmail.com (C.-H.H.); whm526@adm.cgmh.org.tw (H.-M.W.) 3 Department of Chemical and Materials Engineering, Chang Gung University, Taoyuan City 33302, Taiwan; b74225@hotmail.com 1 Graduate Institute of Biomedical Engineering, Chang Gung University, Taoyuan City 33302, Taiwan; l329735@ms49.hinet.net (C.-J.L.); maple7530@gmail.com (Y.-X.Z.); fjiun@hotmail.com (F.-C.H.); d94522010@ntu.edu.tw (W.-P.C.) 4 Department of Chemical Engineering, Ming Chi University of Technology, New Taipei City 24301, Taiwan |
AuthorAffiliation_xml | – name: 3 Department of Chemical and Materials Engineering, Chang Gung University, Taoyuan City 33302, Taiwan; b74225@hotmail.com – name: 1 Graduate Institute of Biomedical Engineering, Chang Gung University, Taoyuan City 33302, Taiwan; l329735@ms49.hinet.net (C.-J.L.); maple7530@gmail.com (Y.-X.Z.); fjiun@hotmail.com (F.-C.H.); d94522010@ntu.edu.tw (W.-P.C.) – name: 2 Division of Haematology/Oncology, Department of Internal Medicine, Chang Gung Memorial Hospital (Linko), Taoyuan City 33302, Taiwan; wisdom5000@gmail.com (C.-H.H.); whm526@adm.cgmh.org.tw (H.-M.W.) – name: 4 Department of Chemical Engineering, Ming Chi University of Technology, New Taipei City 24301, Taiwan |
Author_xml | – sequence: 1 givenname: Chia-Jung surname: Liao fullname: Liao, Chia-Jung – sequence: 2 givenname: Chia-Hsun surname: Hsieh fullname: Hsieh, Chia-Hsun – sequence: 3 givenname: Tzu-Keng surname: Chiu fullname: Chiu, Tzu-Keng – sequence: 4 givenname: Yu-Xian surname: Zhu fullname: Zhu, Yu-Xian – sequence: 5 givenname: Hung-Ming surname: Wang fullname: Wang, Hung-Ming – sequence: 6 givenname: Feng-Chun surname: Hung fullname: Hung, Feng-Chun – sequence: 7 givenname: Wen-Pin surname: Chou fullname: Chou, Wen-Pin – sequence: 8 givenname: Min-Hsien surname: Wu fullname: Wu, Min-Hsien |
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SubjectTerms | Antigens Blood circulation Cancer cancer metastasis Cell adhesion cell isolation circulating tumour cells (CTCs) Clinical significance Design Dielectrophoresis Epithelium Fluorescence Gene expression Glass substrates Hydrogenation Leukocytes Metastasis microfluidic systems Microfluidics optically induced dielectrophoresis (ODEP) Purity |
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Title | An Optically Induced Dielectrophoresis (ODEP)-Based Microfluidic System for the Isolation of High-Purity CD45neg/EpCAMneg Cells from the Blood Samples of Cancer Patients—Demonstration and Initial Exploration of the Clinical Significance of These Cells |
URI | https://www.proquest.com/docview/2582831399 https://search.proquest.com/docview/2179519061 https://pubmed.ncbi.nlm.nih.gov/PMC6266761 https://doaj.org/article/ecaaaf93a9fd49cf890e0447a7248de9 |
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