Capture and release of viable CTCs in microfluidic channel

2017-10-04
Ateş, Hatice Ceren
Şen Doğan, Begüm
Özgür, Ebru
Külah, Haluk
The number of circulating tumor cells (CTCs) in blood is associated with prognosis in several types of cancer. Isolation and characterization of CTCs have important clinical significance in terms of prognosis and early detection of response to treatment. Moreover, downstream characterization of CTCs may help better patient stratification and therapy guidance. However, CTCs are extremely rare and highly sensitive and specific technology is required to isolate viable CTCs from blood cells. In this study, a surface modification strategy is developed for (i) on-chip detection of CTCs by using immunostaining and (ii) viable cell release for downstream analysis. In the first application, selective CTC capture is demonstrated using breast cancer cells (MCF-7) spiked in buffer containing background leukocytes with cell concentration ratio of 1:100. Cells captured by anti-EpCAM coated microchannel are stained with immunofluorescence conjugated anti-pan cytokeratin antibody for epithelial cells and anti-CD45 antibody for hematologic cells, and by DAPI for nuclear staining. Cytokeratin and DAPI stained cells are scored as CTCs whereas CD45+ cells are scored as leukocytes. In the second application, captured cells are released with the combination of enzymatic treatment and high flow rate washing. The viability of released cells is confirmed by observing cell growth in culture.

Suggestions

Capture of circulating tumor cells from blood on modified gold surfaces inside the microfluidic channels
Çetin, Didem; Külah, Haluk; Department of Biomedical Engineering (2019)
Detection of circulating tumor cells (CTCs) from the bloodstream has a critical role in diagnosing and treatment of cancer. However, the number of CTCs in blood compared to other blood cells are extremely rare. In this thesis, various surface modifications strategies for detection of CTCs are studied in order to be used in the microfluidic detection systems. Functionalizing the gold surface with Self Assembled Monolayers (SAMs) used for attaching the EpCAM antibodies, which made possible to immobilize EpCAM...
Capture of rare circulating tumor cells from blood on bio-activated oxide surface inside microfluidic channels
Ateş, Hatice Ceren; Külah, Haluk; Özgür, Ebru; Department of Micro and Nanotechnology (2018)
Isolation and characterization of circulating tumor cells (CTCs) have important clinical significance in terms of prognosis and early detection of response to treatment. Moreover, downstream characterization of CTCs may help better patient stratification and therapy guidance. However, CTCs are extremely rare (~10 CTCs/1010 peripheral blood cells) and highly sensitive, and specific technology is required for their isolation. Rapidly developing microfluidic technologies offer variety of advantages in rare cel...
Enrichment of MCF7 breast cancer cells from leukocytes through continuous flow dielectrophoresis
Çağlayan, Zeynep; Külah, Haluk; Department of Electrical and Electronics Engineering (2018)
Circulating tumor cells (CTCs) are cancerous cells detached from a primary tumor site and enter the bloodstream, causing the development of new tumors in a secondary site. Therefore, their detection in blood is critical to assess the metastatic progression and to guide the line of the therapy. However, the rarity of CTCs in the bloodstream and the lack of suitable detection tool hinders their use as a biomarker in malignancies. Recent advances in microfluidic technologies enabled development of point-of-car...
Synthesis of novel indole-isoxazole hybrids and evaluation of their cytotoxic activities on hepatocellular carcinoma cell lines
Hawash, Mohammed; Kahraman, Deniz Cansen; Ergun, Sezen Guntekin; Cetin-Atalay, Rengul; BAYTAŞ, SULTAN (2021-12-01)
Background Liver cancer is predicted to be the sixth most diagnosed cancer globally and fourth leading cause of cancer deaths. In this study, a series of indole-3-isoxazole-5-carboxamide derivatives were designed, synthesized, and evaluated for their anticancer activities. The chemical structures of these of final compounds and intermediates were characterized by using IR, HRMS, H-1-NMR and C-13-NMR spectroscopy and element analysis. Results The cytotoxic activity was performed against Huh7, MCF7 and HCT116...
Examination of the dielectrophoretic spectra of MCF7 breast cancer cells and leukocytes
Çağlayan, Zeynep; Demircan Yalçın, Yağmur; Külah, Haluk (Wiley, 2020-03-01)
The detection of circulating tumor cells (CTCs) in blood is crucial to assess metastatic progression and to guide therapy. Dielectrophoresis (DEP) is a powerful cell surface marker-free method that allows intrinsic dielectric properties of suspended cells to be exploited for CTC enrichment/isolation from blood. Design of a successful DEP-based CTC enrichment/isolation system requires that the DEP response of the targeted particles should accurately be known. This paper presents a DEP spectrum method to inve...
Citation Formats
H. C. Ateş, B. Şen Doğan, E. Özgür, and H. Külah, “Capture and release of viable CTCs in microfluidic channel,” presented at the 9th Annual Lab-on-a-Chip and Microfluidics World Congress, 2017, California, USA, 2017, Accessed: 00, 2021. [Online]. Available: https://hdl.handle.net/11511/86857.