Al Nasser, M. (Maryem) (2013) Multiple approaches to image cell death. Study in an in vitro model of cisplatin-sensitive human testicular carcinoma. thesis, Medicine.
Full text available on request.Abstract
Introduction: Apoptosis (programmed cell death) is an important determinant of tumour response to anticancer agents. Imaging of cell death is therefore crucial in diagnosing diseases at early stages and in monitoring treatment. Using non-invasive techniques that detect early cell death is very important. This can be achieved by using functional molecular imaging such as PET. In this study multiple probes are pre-clinically evaluated for their ability in cell death detection. Materials and methods: Apoptosis was induced in TERA cells treated with increasing doses of cisplatin. Several in vitro techniques (fluorescent microscopy, flow cytometry, radioligand binding assay) were used to measure cell death. FITC-labelled Annexin A5 and AF546- and AF647-labelled Annexin A5 were used to detect phosphatidylserine exposure in apoptotic cell using fluorescent microscopy and flow cytometry. For fluorescent microscopy, DAPI was used to detect later apoptotic cells or necrotic cells. For flow cytometry, Annexin A5 staining was compared with DiOC6 (changes in mitochondrial membrane potential) and with propidium iodide (increase in cell membrane permeability). Furthermore, Annexin A5 staining was combined with propidium iodide to discriminate between cell death modes. Radiolabeled AnnexinA5 (99mTc-HIS-cys-AnxA5) was also used. Results: Annexin A5 can detect early and late apoptotic cells by binding to the phosphatidylserine. Labelled Annexin A5 proved to be specific in its binding to phosphatidylserine and in its detection of only apoptotic cells. PI can detect the late apoptotic and necrotic cells with increased plasma membrane permeability. DiOC6 detects decrease in mitochondrial membrane potential, which occurs early after onset of apoptosis. Conclusions: Different probes, such as fluorescently and radio-labeled Annexin A5 and DiOC6 are useful methods for apoptosis imaging. However, radiolabeled Annexin A5 (99mTc-HIS-cys-AnxA5) may be a promising tool for cell death imaging in vivo.
| Item Type: | Thesis (Thesis) |
|---|---|
| Supervisor name: | Boersma, Dr. Hendrikus H. and Rybczynska, Dr. Anna A. |
| Faculty: | Medical Sciences |
| Date Deposited: | 25 Jun 2020 10:43 |
| Last Modified: | 25 Jun 2020 10:43 |
| URI: | https://umcg.studenttheses.ub.rug.nl/id/eprint/508 |
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