Medizinische Universität Graz - Research portal
Selected Publication:
Hoegler, A.
In vitro characterization of adipose-derived mesenchymal stromal cells and comparison of PCL/PLA scaffolds and Matriderm for their suitability as cell-seeded grafts.
[ Diplomarbeit/Master Thesis (UNI) ] Universität Graz; 2022.
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- Authors Med Uni Graz:
- Advisor:
- Lang-Olip Ingrid
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- Abstract:
- Adipose-derived mesenchymal stromal cells (AD-MSCs) are a great source for cell-based applications, especially in regenerative medicine. A special focus was placed on skin substitutes such as polycaprolactone/polylactide or Matriderm as carriers for stem cell applications. The aim of the study was to isolate and characterize AD-MSCs from 9 different probands in vitro to select cells of the best donor for their further application in vivo experiments. In addition, PCL/PLA scaffolds and Matriderm were compared to determine the carrier with the best cell-seeding conditions. AD-MSCs were isolated from the stromal vascular fraction and characterized by immunohistochemistry and flow cytometry. The data revealed that cells were positive for CD90, CD73 and CD105 and mainly lacked the expression of CD45, CD34 and HLA-DR. Immunohistochemical staining showed that cells of donor 3, 4 and 5 were not suitable for further experiments due to smA contamination and high CD45 expression. Cells of proband 2, 6, 7, 8, 9 and 10 showed a good morphology and were well suited for ongoing studies. Based on immunohistochemistry and flow cytometry a homogenous cell population can be observed from the third passage onwards. Both flow cytometry and proliferation assay affirmed that the cells should only be used up to passage 7, since viability then decreased. AD-MSCs were cultured in adipogenic and osteogenic medium for 14 days and 21 days, respectively to evaluate their differentiation potential. Adipogenic induction led to accumulation of lipid droplets and calcium deposits were visible by osteogenic differentiation. In addition, osteogenic-induced AD-MSCs showed a strikingly enhanced formation of lipid droplets. Ultrastructural and histological evaluations revealed that PCL/PLA and Matriderm offered good cell-culture conditions for AD-MSCs, which makes them suitable for their application in continuing wound healing studies.