Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Fuchs, J.
Flexibility in Organ Reserach - A 3D Cell Culture Model of the Human Placenta.
Doktoratsstudium der Medizinischen Wissenschaft; Humanmedizin; [ Dissertation ] Graz Medical University; 2020. pp. 96
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Brislinger Dagmar
- Lang-Olip Ingrid
- Stadlbauer-Köllner Vanessa
- Altmetrics:
- Abstract:
- Trophoblast invasion into the maternal endovascular system is a crucial step in the implantation process of the human blastocyst. Failures in this process are often associated with pregnancy pathologies that cause severe complications for the mother and the fetus. Therefore, it is necessary to establish innovative research methods to improve our knowledge about placental implantation and enable new kinds of treatments. 3D cell culture systems play a major role in the investigation of these processes, as they offer the possibility to reflect in vivo-like situations. A novel matrix-based 3D placenta cell culture system to investigate the interaction of trophoblasts and maternal endothelial cells in early human pregnancy is currently established in our lab. This system uses biodegradable and thermosensitive PCL/PLA hollow fibers. An appropriate histological processing method and validation of antibodies for the characterization of placenta specific cell types are basic requirements to fulfil the needs of this 3D placenta model. The thermosensitive PCL/PLA samples were histological processed with our established low-melting-point paraffin embedding method focusing on the histological examination of the trophoblast migration towards endothelial cells under normoxic (21% O2) and hypoxic (2.5% O2) conditions. By the combination of a low-melting-point paraffin embedding and a heat-reduced enzymatic antigen retrieval method it was possible to identify and analyse migrated cells within the membrane. Computational assisted evaluation was done to analyse the samples in a quantitative and qualitative way. Quantitative analyses of the immunofluorescent staining showed the intensity pattern of 17 placenta specific antibodies. Qualitative analysis revealed that certain antigen retrieval methods lead to the staining of several cell types within the tissue. The successful establishment of low-melting-point paraffin embedding combined with enzymatic antigen retrieval with pepsin for the histological characterization, revealed proliferation of trophoblast cells under normoxic conditions whereas cells started to migrate into PCL/PLA fiber network of the membrane under hypoxic conditions in our novel 3D placental cell culture system. Using the cell line ACH-3P, trophoblast migration towards primary endothelial cells also was shown under co-culture conditions. In addition, it is important to mention that for histological preparations a quantitative analysis must be accompanied by a qualitative analysis and vice versa in order to provide a comprehensive interpretation of the data.