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Title: Negative Enrichment of Circulating Tumor Cells Via 3D Printed Microfluidic Device
Committee:
Dr. Fatih Sarioglu, ECE, Chair , Advisor
Dr. Albert Frazier, ECE
Dr. Stanislav Emelianov, ECE
Dr. Levent Degertekin, ECE
Dr. John McDonald, Biological Sciences
Abstract: The enrichment of CTCs from the blood samples is important for clinical management of cancer and understanding the biology of the cancer metastatic process but difficult due to the rarity and heterogeneity of CTCs. Compared to conventional benchtop approaches, such as density gradient centrifugation and scanning cytometry, microfluidics approaches offer higher sensitivity and specificity with deterministically screening of CTC in a controlled microenvironment. For label-free and unbiased enrichment of CTCs in a microfluidic device, negative enrichment technique that removes the leukocytes through antigen-antibody interaction is commonly employed but suffers from several technological hurdles. First, limited surface area on the conventional microfluidic device leads to surface saturation due to a large number of leukocytes in clinically relevant volumes of blood sample. Second, sample preparation, such as pre-lysing of the red blood cells, can drastically increase the sample volume while prone to losing valuable tumor cells. In order to address these shortcomings, I developed a 3D printed microfluidic device that integrates stacked layers of chemically functionalized leukodepletion microfluidic channels and portable microfiltration compartment in a monolithic device to provide high throughput and label-free enrichment of CTC. The stacked leukodepletion channels provide sufficient surface area to accommodate the leukocytes in clinically relevant volumes of the blood sample while the post-microfiltration efficiently removes the red blood cells from the enriched blood sample and retains the CTCs on the membrane filter. The portability of the membrane filter allows for the downstream analysis of the recovered CTCs with a microscope or micromanipulator. With the devices that I have developed, I was able to process 10 mL of whole blood samples and recover CTCs from prostate and pancreatic cancer patients' blood samples.
