Optimizing the isolation and storage conditions of mitochondria from human umbilical cord stroma mesenchymal stem cells

In recent years, it has been reported that mitochondria can be transferred between cells in a natural process and as a result, advantages such as intercellular energy synchronization are provided. Ongoing research on the subject has shown that mitochondria transfer can be performed artificially under both in vitro and in vivo conditions. Mitotherapy, ultimately emerging as a new therapeutic approach, is based on the successful transplantation of healthy exogenous mitochondria for the treatment of diseases related to damage and dysfunction of mitochondria. It is important that the mitochondria used at this stage are isolated from healthy cells and maintained under optimized conditions until transplantation. In this thesis study, it was preferred to use Mesenchymal Stem Cells (MSCs) to provide a good source for mitochondrial isolation and also to have regenerative properties in various studies. After the mitochondria of human umbilical cord stromal MSCs (WJ-MSCs) were isolated, they were frozen and preserved for the first time by the Vitrification method, which is suitable for cryopreservation of small-volume samples. The mtDNA copy number of mitochondria before and after cryopreservation was calculated quantitatively by qPCR. In addition, as a result of the transfer of fresh and frozen mitochondria to WJ-MSCs, their functionality was evaluated according to cell proliferation and correlated with the results of other analyses.