Combination of antibody-coated, physical-based microfluidic chip with wave-shaped arrays for isolating circulating tumor cells
- 375 Downloads
Circulating tumor cells (CTCs) are found in the peripheral blood of patients with metastatic cancers, which have critical significance in cancer prognosis and diagnostics. Enumeration is significantly valuable since number of CTCs is strongly correlated to severity of disease. This article is proposed and demonstrated an antibody-coated, size-based microfluidic chip with wave-shaped arrays could efficiently capture CTCs combining two separation methods of both size- and deformability-based and affinity-based segregation. Utilizing immunocapture of capture chemistry of Epithelial Cell Adhension Molecule (EpCAM), tumor cells could be captured by narrow gaps or have a friction with microposts edges to realize both immune-affinity and size capture. This wave-shaped layout of microfluidic chip with varying gaps between adjacent circular microposts can generate perpendicular velocities to the fluidic direction. This oriented fluidic direction will carry cells to next smaller neighboring gap and then be captured gradually. The experiment results indicate capture efficiency is ~90% and viability is ~95% after extracted and cultured 3 days. Furthermore, this chip has been validated for whole blood with cancer cell lines and mimic patient blood. This study demonstrates feasibility using our microfluidic chip for CTCs research, monitoring cancer progress and evaluating therapeutic treatment.
KeywordsCirculating tumor cells (CTCs) Epithelial cell adhension molecule (EpCAM) Epithelial-to-mesenchymal transition (EMT)
This research work was supported by the Major State Basic Research Development Program of China (973 Program) (Grant No. 2011CB933102), National Natural Science Foundation of China (Key Program, Grant No. 61335010), Key Research Program of Chinese Academy of Sciences (Grant No. KJZD-EW-TZ-L03-6) and Postdoctoral Research Funding (Grant No.2014 M550794). We deeply appreciate Prof. Yu-Sheng Lin from Sun Yat-Sen University helped with editing and revising the manuscript. Jingjing zhang from Xi’ an Technological University helped with simulation. Prof. Fan from Florida University, Prof. Chen from École Normale Supérieure (ENS) and Prof. Yang from Chinese Academy of Sciences offered deep discussion. Zhaoxin Geng, Graduate student Hongsheng Gao and Xiaoqing Lv from Chinese Academy of Sciences helped with design and modification, respectively. Graduate student Zhili Wang from Chinese Academy of Sciences helped to culture cells. Barbara Costello offered help for revising the manuscript.
- A.A. Adams, P.I. Okagbare, J. Feng, M.L. Hupert, D. Patterson, J. Gottert, R.L. McCarley, D. Nikitopoulos, M.C. Murphy, S.A. Soper, Highly efficient circulating tumor cell isolation from whole blood and label-free enumeration using polymer-based microfluidics with an integrated conductivity sensor. J. Am. Chem. Soc. 130, 8633–8641 (2008)CrossRefGoogle Scholar
- I. Desitter, B.S. Guerrouahen, N. Benali-Furet, J. Wechsler, P.A. Janne, Y. Kuang, M. Tanagita, L. Wang, J.A. Berkowitz, R.J. Distel, Y.E. Cayre, A new device for rapid isolation by size and characterization of rare circulating tumor cells. Anticancer Res. 31, 427–441 (2011)Google Scholar
- J.P. Gleghorn, E.D. Pratt, D. Denning, H. Liu, N.H. Bander, S. Tagawa, D.M. Nanus, P.A. Giannakakou, B.J. Kirby, Capture of circulating tumor cells from whole blood of prostate cancer patients using geometrically enhanced differential immunocapture and a prostate-specific antibody. Lab Chip 10, 27–29 (2010)CrossRefGoogle Scholar
- N.M. Karabacak, P.S. Spuhler, F. Fachin, E.J. Lim, V. Pai, E. Ozkumur, J.M. Martel, N. Kojic, K. Smith, P. Chen, J. Yang, H. Hwang, B. Morgan, J. Trautwein, T.A. Barber, S.L. Stott, S. Maheswaran, R. Kapur, A.A. Haber, M. Toner, Microfluidic, marker-free isolation of circulating tumor cells from blood samples. Nature 9 (2014)Google Scholar
- P. Katharina, C. Oumar, K. Andreas, K. Sabine, M. Nele, G. Mieczyslaw, K. Torsten, J. Cornelia, H. Ulrike, A.H. Annelore, R. Carola, P. Ulrich, R. Ingo, H. Klaus, Monitoring the response of circulating epithelial tumor cells to adjuvant chemotherapy in breast cancer allows detection of patients at risk of early relapse. J. Clin. Oncol. 26, 1208–1215 (2008)CrossRefGoogle Scholar
- S. Nagrath, L.V. Sequist, S. Maheswaran, D.W. Bell, D. Irimia, L. Ulkus, M.R. Smith, E.L. Kwak, S. Digumarthy, A. Muzikansky, P. Ryan, U.J. Balis, R.G. Tompkins, D.A. Haber, M. Toner, Isolation of rare circulating tumour cells in cancer patients by microchip technology. Nature 450, 1235–1239 (2007)CrossRefGoogle Scholar
- X. Qin, S. Park, S.P. Duffy, K. Matthews, R.R. Ang, T. Todenhöfer, H. Abdi, A. Azad, J. Bazov, K.N. Chi, P.C. Black, H. Ma, Size and deformability based separation of circulating tumor cells from castrate resistant prostate cancer patients using resettable cell traps. Lab Chip 15, 2278–2286 (2015)CrossRefGoogle Scholar
- M. Shyamala, V.S. Lecia, N. Sunitha, U. Lindsey, B. Brian, V.C. Chey, I. Elizabeth, D. Sven, A. JI, W.B. Daphne, D. Subba, M. Alona, I. Daniel, S. Jeffrey, G.T. Ronald, J.L. Thomas, T. Mehmet, A.H. Daniel, Detection of mutations in EGFR in circulating lung-cancer cells. N. Engl. J. Med. 359, 366–377 (2008)CrossRefGoogle Scholar
- S.L. Stott, C.H. Hsu, D.I. Tsukrov, M. Yu, D.T. Miyamoto, B.A. Waltman, S.M. Rothenberg, A.M. Shah, M.E. Smas, G.K. Korir, F.P. Floyd, A.J. Gilman Jr., J.B. Lord, D. Winokur, S. Springer, D. Irimia, S. Nagrath, L.V. Sequist, R.J. Lee, K.J. Isselbacher, S. Maheswaran, D.A. Haber, M. Toner, Isolation of circulating tumor cells using a microvortex-generating herringbone-chip. Proc. Natl. Acad. Sci. U. S. A. 107, 18392–18397 (2010)CrossRefGoogle Scholar
- S. Wang, K. Liu, J. Liu, Y.K. Lee, Z.T.F. Yu, E.K. Lee, X. Xu, J. Reiss, L.W.K. Chung, J. Huang, M. Rettig, A. Pantuck, D. Silegson, K.N. Duraiswamy, C.K.F. Shen, H.R. Tseng, Highly efficient capture of circulating tumor cells using nanostructured silicon substrates with integrated chaotic micromixers. Angew. Chem. Int. Ed. 50, 3084–3088 (2011)CrossRefGoogle Scholar