Donor tissues from cadavers represent a valuable alternative source for the isolation of primary cells with mesenchymal stem/stromal cells (MSC)-like properties. However, the properties of primary cells obtained from different tissues and at different time points after death are poorly understood. In this doctoral thesis, we aimed to identify the optimal tissue source among three knee and peri-knee tissues for the isolation of primary cells with MSC-like properties, and to determine the influence of time after death on the properties of these cells. Tissues from the subchondral bone, synovium and periosteum were harvested from 32 donors at various post mortem intervals. Primary cells were analysed using routine in vitro methods such as colony-forming unit fibroblast assay, trilinear differentiation, immunophenotyping, and novel analyses, such as gene expression of recently identified markers of human skeletal stem cells. Our results show that primary cells isolated from three tissues are similar in MSC-specific properties. However, we detected increased expression of the gene encoding negative marker of human skeletal stem cells, CD146, in bone marrow cells. We also showed that the success rate of primary cell isolation depends on the post mortem interval. However, we also found that cells from the synovium and periosteum isolated more than 48 hours after death had better osteogenic and chondrogenic potential compared to cells from the same tissues isolated faster after death. The results of this thesis demonstrate that knee and peri-knee tissues of cadaveric donors are a strategic source of primary cells with MSC-like properties that can be used in regenerative medicine as even three days after death.
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