University of Twente Student Theses


Characterization of circulating tumour cells in prostate cancer: viability and secretion studies after enrichment with EpCAM targeting ferrofluids

Bante, Gidon (2022) Characterization of circulating tumour cells in prostate cancer: viability and secretion studies after enrichment with EpCAM targeting ferrofluids.

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Abstract:Prostate cancer (PCa) is the most common cancer among men in the USA and the second most death causing cancer when looking at total numbers. Primary PCa has nowadays 5 year survival rate of 100%, however metastatic PCa only has a 32% 5 year survival rate. Different PCa types need different treatment depending the stage of the cancer, AR dependency, aggressiveness, and mutations. That causes many prescribed treatments to be unresponsive or only partly responsive. More personalised treatment is needed which counters the huge heterogeneity issue of PCa. We believe that CTCs are the key to understanding a patients exact disease profile. Therefore we developed a method to look at these CTCs individually. The biggest problem with CTC is their extreme rarity, there is maybe 1 CTC in a 7.5 mL blood sample. By use of DLA, immunomagnetic enrichment, and a microwell chip which causes cell isolation we tackle this needle in a haystack problem. In this research we look at if the use of immunomagnetic ferrofluids (FF) for enrichment influences the CTCs. With Live/Dead stainings and flowcytometry we determined that there is some cell dead among CTC caused by FF. Next to that, metabolic activity was measured with Alamar Blue Assays which pointed out that metabolic activity did not decrease. Which is counter-intuitive with the lower viability results. Cell death was not on a level that it would make the single CTC analysis unworkable. Chips from Vycap were used to make the first start towards single cell secretion analysis where patterns of Prostate Specific Antigen (PSA) were found. Optimization of current protocols will likely result in images that can be analysed and compared. We also observed that there was no visual difference between cells and enriched cells (with FF). Again, with optimization this can give a platform for analysis and comparison. Lastly we worked with PCa patient DLA samples in order to catch CTC on the Vycap chip. This proved to be difficult and even though some CTCs were found, secretion was not seen. In order to improve the CTC numbers that were found, we recommend using an extra enrichment method to filter out even more cell debris and leukoctyes of the DLA sample. That will open the future to secretion analysis of single CTC. Future work with PCa cells will make it possible to ’punch out’ these cells of the chip and culture them for medicine testing. Which will lead to personalised medicine, better treatment decisions and more survivors of metastatic prostate cancer.
Item Type:Essay (Master)
Faculty:TNW: Science and Technology
Subject:30 exact sciences in general, 42 biology, 44 medicine
Programme:Biomedical Engineering MSc (66226)
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