Dokument: Two magnetic enrichment methods for pancreatic circulating tumour cells detection
| Titel: | Two magnetic enrichment methods for pancreatic circulating tumour cells detection | |||||||
| URL für Lesezeichen: | https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=62400 | |||||||
| URN (NBN): | urn:nbn:de:hbz:061-20230419-105148-8 | |||||||
| Kollektion: | Dissertationen | |||||||
| Sprache: | Englisch | |||||||
| Dokumententyp: | Wissenschaftliche Abschlussarbeiten » Dissertation | |||||||
| Medientyp: | Text | |||||||
| Autor: | Wu, Junhao [Autor] | |||||||
| Dateien: |
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| Beitragende: | Prof.Dr. Stoecklein, Nikolas [Gutachter] Prof. Dr. rer. nat. Neubauer, Hans [Gutachter] | |||||||
| Dewey Dezimal-Klassifikation: | 600 Technik, Medizin, angewandte Wissenschaften » 610 Medizin und Gesundheit | |||||||
| Beschreibung: | In the last decades there has been significant technical developments in the field of liquid biopsies that have improved our access to circulating tumour cells (CTCs). However, for some tumour types, as the pancreatic ductal adenocarcinoma, the detection rate and number of CTCs detected are still very low which compromises the use of CTCs as biomarker and tumour tissue surrogate for molecular analysis. A major limitation resides on the low volume of blood that is screened in standard CTC assays. Diagnostic leukaphaeresis allow to sample liters of blood and constitutes a promising alternative strategy to collect higher numbers of CTCs, however the high cellular concentration of these products constitutes additional challenging for CTC enrichment with the existing CTC assays. In the present work I explored the technical performance of two systems for enrichment and detection of rare tumour cells in highly concentrated blood products: The Isoflux and the KingFisher. The first is commercially available specifically for CTC enrichment purposes, while the second system required the design and optimization of protocols for this application. Both methods are based in immunomagnetic enrichment and I tested the performance using several types of magnetic beads and using EpCAM and MUC-1 as epitopes targeted at surface of the tumour cells. Despite enrichment of rare cells was possible with both systems, in the particular experimental setting, the performance of KingFisher was superior. Furthermore, in the KingFisher I could unify enrichment and subsequent immunostaining in a single automated protocol reducing the hands-on time necessary. Additionally, in the KingFisher system, I could establish more complex protocols combining different beads for initial depletion of hematogenous cells prior positive selection of CTCs. This work demonstrates the high flexibility in terms of magnetic beads and the high cost-effectiveness. It also shows the possibility to automate staining and reduce the operator intervention, and the possibility to parallel process a high number of samples in the KingFisher system. The results collected suggest that it might constitute an attractive solution to process large blood volumes. | |||||||
| Lizenz: |  Dieses Werk ist lizenziert unter einer Creative Commons Namensnennung 4.0 International Lizenz | |||||||
| Fachbereich / Einrichtung: | Zentrale Einrichtungen » Universitäts- und Landesbibliothek (ULB) | |||||||
| Dokument erstellt am: | 19.04.2023 | |||||||
| Dateien geändert am: | 19.04.2023 | |||||||
| Promotionsantrag am: | 27.10.2022 | |||||||
| Datum der Promotion: | 11.04.2023 |
