Medizinische Universität Graz - Research portal
Selected Publication:
Kinzer, M.
In vivo effects of fibroblasts and mesenchymal stromal cells on angiogenic processes in a mouse model.
[ Diplomarbeit/Master Thesis ] Karl-Franzens University Graz; 2013.
- Authors Med Uni Graz:
- Advisor:
- Lang-Olip Ingrid
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- Abstract:
- Abstract Cell transplantation of mesenchymal stromal cells (MSC) is considered as a promising strategy for the revascularization of ischemic tissue. In this context, MSC from the human placenta are attractive candidates. They are not only available in large supply but can further reduce rejection and inflammation due to their low immunogenicity. Recently, it was shown that both amnion-derived MSC (hAMSC) as well as placental fibroblasts (PlFb1), isolated from chorionic vessel explants, enhance endothelial cell viability in vitro via secretion of angiogenic factors. The aim of this study was to evaluate the angiogenic effects of hAMSC and P1Fb1 in vivo. In addition, the phenotype of the cells should be compared. Both cell types were isolated from human term placentas and characterized by flow cytometry and immunocytochemistry. For in vivo experiments, cells were suspended in Matrigel and injected subcutaneously into the flank of immune-deficient mice (NOG mice). hAMSC and PlFb1 were applied both separately or combined with placental endothelial cells (PlEC). Matrigel without cells or containing PlEC served as the respective negative or positive control. Matrigel plugs were excised after 14 or 28 days and analyzed histologically. Subsequently, blood vessels of human origin were quantified. The two cell types, namely hAMSC and PlFbl, showed no difference in the expression of various surface markers (CD10+, CD13+, CD29+, CD49a+, CD63+, CD73+, CD90+, CD105+, CD166+, HLA-ABC+, CD3-, CD14-, CD15-, CD19-, CD31-, CD45-, CD271-, HLA-DR-, AP-, MSCA-1-, myosin-, desmin-, smActin-). hAMSC alone did not induce any capillary formation in vivo, but prevented immigration of mouse cells into the plug. Application of P1EC did not show this effect, yet formed few human capillaries, which covered up to 0.02 % of the Matrigel area. The highest density of capillaries was achieved by the combined application of PlEC plus hAMSC, covering 0.27 1.44%. Immunohistochemistry with specific antibodies confirmed the successful connection of the newly formed human capillaries to the mouse vasculature. In vivo experiments with PlFb1 did not lead to representative results, due to a reduced functionality of the applied PlEC. In summary, it was shown that hAMSC and P1Fb1 are similar regarding morphology and expression of numerous surface markers. In vivo studies indicate pro-angiogenic effects of hAMSC. Therefore, the angiogenic effects of P1Fb1 should be further investigated to gain insight between similarities and differences of hAMSC and P1F1b with the aim to find attractive candidates for cell-based applications in vascular medicine.