Isolation of circulating tumor cells using a microvortex-generating herringbone-chip
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 capturin...
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Published in | Proceedings of the National Academy of Sciences - PNAS Vol. 107; no. 43; pp. 18392 - 18397 |
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Main Authors | , , , , , , , , , , , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
National Academy of Sciences
26.10.2010
National Acad Sciences |
Subjects | |
Online Access | Get full text |
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Abstract | 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. |
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AbstractList | 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. 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. [PUBLICATION ABSTRACT] 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. |
Author | Gilman, Anna J. Shah, Ajay M. Nagrath, Sunitha Winokur, Daniel Irimia, Daniel Smas, Malgorzata E. Lord, Jenna B. Waltman, Belinda A. Lee, Richard J. Yu, Min Miyamoto, David T. Tsukrov, Dina I. Isselbacher, Kurt J. Maheswaran, Shyamala Haber, Daniel A. Stott, Shannon L. Floyd, Frederick P. Springer, Simeon Hsu, Chia-Hsien Korir, George K. Rothenberg, S. Michael Sequist, Lecia V. Toner, Mehmet |
Author_xml | – sequence: 1 givenname: Shannon L. surname: Stott fullname: Stott, Shannon L. – sequence: 2 givenname: Chia-Hsien surname: Hsu fullname: Hsu, Chia-Hsien – sequence: 3 givenname: Dina I. surname: Tsukrov fullname: Tsukrov, Dina I. – sequence: 4 givenname: Min surname: Yu fullname: Yu, Min – sequence: 5 givenname: David T. surname: Miyamoto fullname: Miyamoto, David T. – sequence: 6 givenname: Belinda A. surname: Waltman fullname: Waltman, Belinda A. – sequence: 7 givenname: S. Michael surname: Rothenberg fullname: Rothenberg, S. Michael – sequence: 8 givenname: Ajay M. surname: Shah fullname: Shah, Ajay M. – sequence: 9 givenname: Malgorzata E. surname: Smas fullname: Smas, Malgorzata E. – sequence: 10 givenname: George K. surname: Korir fullname: Korir, George K. – sequence: 11 givenname: Frederick P. surname: Floyd fullname: Floyd, Frederick P. – sequence: 12 givenname: Anna J. surname: Gilman fullname: Gilman, Anna J. – sequence: 13 givenname: Jenna B. surname: Lord fullname: Lord, Jenna B. – sequence: 14 givenname: Daniel surname: Winokur fullname: Winokur, Daniel – sequence: 15 givenname: Simeon surname: Springer fullname: Springer, Simeon – sequence: 16 givenname: Daniel surname: Irimia fullname: Irimia, Daniel – sequence: 17 givenname: Sunitha surname: Nagrath fullname: Nagrath, Sunitha – sequence: 18 givenname: Lecia V. surname: Sequist fullname: Sequist, Lecia V. – sequence: 19 givenname: Richard J. surname: Lee fullname: Lee, Richard J. – sequence: 20 givenname: Kurt J. surname: Isselbacher fullname: Isselbacher, Kurt J. – sequence: 21 givenname: Shyamala surname: Maheswaran fullname: Maheswaran, Shyamala – sequence: 22 givenname: Daniel A. surname: Haber fullname: Haber, Daniel A. – sequence: 23 givenname: Mehmet surname: Toner fullname: Toner, Mehmet |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/20930119$$D View this record in MEDLINE/PubMed |
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Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 3Present address: Division of Medical Engineering Research, National Health Research Institutes, No. 35, Keyan Road, Zhunan Town, Miaoli County, 35053, Taiwan. Contributed by Kurt J. Isselbacher, August 24, 2010 (sent for review July 14, 2010) Author contributions: S.L.S., C.-H.H., D.I.T., M.Y., D.T.M., B.A.W., S.M.R., A.M.S., G.K.K., D.I., S.N., L.V.S., R.J.L., K.J.I., S.M., D.A.H., and M.T. designed research; S.L.S., C.-H.H., D.I.T., M.Y., D.T.M., B.A.W., S.M.R., A.M.S., M.E.S., G.K.K., F.P.F., A.J.G., J.B.L., D.W., and S.S. performed research; S.L.S., C.-H.H., D.I.T., M.Y., D.T.M., B.A.W., S.M.R., M.E.S., G.K.K., F.P.F., J.B.L., S.N., R.J.L., K.J.I., S.M., D.A.H., and M.T. analyzed data; and S.L.S., C.-H.H., D.I.T., K.J.I., S.M., D.A.H., and M.T. wrote the paper. 1S.L.S. and C-H.H. contributed equally to this work. |
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