Mesenchymal stem/stromal cells isolated from bone marrow are the most researched and reliable source of stem cells used in regenerative medicine to date. New sources of MSCs, from which they can be obtained more easily compared to bone marrow harvesting, are currently being researched and have shown promising results. Cadaveric donor tissue is also an alternative source of MSCs. The need for a high cell yield for extensive therapies has led to the development of bioreactors – automated cell cultivation systems. Bioreactors allow for better reproducibility and a reduction in labour, time and cost compared to conventional cell cultivation protocols. In this master's thesis, we investigated and compared the osteogenic differentiation capabilities of MSCs isolated from the bone marrow of a young, healthy donor that could be suitable for the production of advanced therapy medicinal products. The cells were cultured in two different ways: manually (labelled RG) and in a cell bioreactor (labelled QB). We compared their osteogenic differentiation potential with cells obtained post mortem from the periosteum of a healthy donor of similar age. We assessed osteogenic differentiation potential using alizarin red S histology and measuring gene expression of osteogenic genes, namely OC, RUNX2 and COL1A1 using quantitative polymerase chain reaction. Comparison of cells obtained from two different sources showed a more extensive formation of calcium deposits and higher osteogenic gene expression in cells from bone marrow. In summary, bone marrow-derived MSCs have a greater osteogenic differentiation potential than cells isolated from the periosteum. Cells cultured in bioreactors and those cultured manually gave comparable results. In summary, MSCs from the bone marrow of a young donor have a suitable osteogenic differentiation potential and can be further tested for their use in cell therapies.
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