Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Weiss, G.
Parakriner Effekt von mesenchymalen Stammzellen der Plazenta (hAMSC) auf die Funktion von Endothelzellen.
[ Diplomarbeit/Master Thesis ] Karl-Franzens University Graz; 2011.
- Autor*innen der Med Uni Graz:
- Weiss Gregor
- Betreuer*innen:
- Lang-Olip Ingrid
- Altmetrics:
- Abstract:
- Abstract Mesenchymal stem cells (MSC) isolated from the human amniotic membrane (hAMSC) are multipotent cells with the ability to promote angiogenesis. Therefore they represent a potential target in regenerative medicine. The cells are a promising alternative source to bone marrow derived stromal cells as they are isolated non-invasively and are immunologically well tolerated. In this study the effect of hAMSC-conditioned medium (hAMSC-Cdm) on endothelial cells and the secretion of angiogenic factors by hAMSC have been investigated and compared with the impact of conditioned media derived from placental endothelial cells (Plaec) and placental fibroblasts (Plfib). The different conditioned media (Cdm) were prepared by incubating confluent hAMSC, Plaec and Plfib with EGM-2 medium for 48 h under 21% (CdmN°Rwl) and 2% (CdmHYP) oxygen culture conditions. The effects of hAMSC-Cdm, Plaec-Cdm and Plfib-Cdm on endothelial viability and network formation were determined using LDH and Matrigel assay, respectively. For detection of angiogenic proteins, Cdm were analysed by an angiogenesis array kit and further EL1SA applications. Non-conditioned medium EGM-2 was used as control. Plaec cultivated in Cdm released significantly less LDH than control cells kept in non-conditioned medium, which indicates their increased cell viability. hAMSC-Cdm induced the highest reduction of LDH (53.5 ± 26.3% of control, p<0.001), closely followed by Plfib-Cdm (58.9 ± 16.8% of control, p<0.001). Plaec-Cdm in contrast showed weaker effects (70.9 ± 8.3% of control, p<0.00I). In the Matrigel Assay, hAMSC-Cdm and Plaec-Cdm respectively showed only a tendency or no effect in supporting endothelial network formation. In contrast, Plfib-Cdm significantly increased the number of endothelial networks. Angiogenesis array analysis mainly revealed a lower level of pro- and anti-angiogenic factors by hAMSC and Plfib compared to Plaec. ELISA analysis showed that Plaec produced P1GF, PDGF and FGF, and Plfib secreted VEGF, with best effects under 2% oxygen conditions. Interestingly, hAMSC did not produce any of the stated growth factors. In conclusion, VEGF secreted by Plfib seems to be responsible for enhanced endothelial network formation. hAMSC and Plfib showed better results on the cell viability than Plaec. These properties of hAMSC and Plfib indicate possible applications in the therapy of cardiovascular diseases.