Obtaining mesenchymal stem cells from human exfoliated deciduous teeth pulp, characterization with cell surface markers (CD90, CD105, CD45), and evaluation of kisspeptin and neurokinin receptor ((KISS1R, TACR3) gene expressions by real-time PCR (RT-PCR)

This study is a master's thesis project named "Characterization of Human Deciduous Mesenchymal Stem Cells with CD90+, CD105+ CD45- Cell Surface Markers and Investigation of Kisspeptin and Neurokinin Receptors (KISS1, KISS1R, TAC3, TACR3) by RT-PCR". It has been prepared with the support of the Research Projects Coordination Unit. The aim of the presented thesis study is that human exfoliated decidous teeth stem cells can be used as an alternative source for reproductive system diseases such as hypogonadism, since they express kisspeptin, neurokinins and their receptors and these avaible genes play a key role in puberty. It was ensured that the deciduous teeth stem cells, whose exfoliation time had come, were isolated from a 9-year-old male patient in accordance with the permissions obtained from the Çukurova University Ethics Committee. The culturing of decidous tooth stem cells, which were isolated, continued at Cukurova University Biotechnology Research and Application Center. Positive (CD90 and CD105) and negative (CD45) markers were determined for flow cytometry analysis. The mesenchymal stem cell character of decidous teeth stem cells was confirmed by cell surface markers. Deciduous stem cells, which were confirmed to have mesenchymal stem cell characteristics, were allowed to reach passages 3, 4 and 5, and first of all, RNA isolation for each passage and then conversion to cDNA was ensured. Expression of genes (KISS1, KISS1R, TAC3 and TACR3) in decidous teeth stem cells was determined by real-time polymerase chain reaction (RT-PCR). KISS1, KISS1R and TAC3, TACR3 gene expressions were analyzed in RT-PCR for passages 3, 4 and 5, and the change in gene expression was correlated with the number of passages. Kisspeptin-1 encoded by the KISS1 gene, and KISS1R as its ligand, and TAC3, TAC3R genes that affect the KISS1-KISS1R (GPR54) system have been associated with the reproductive system. It has been focused on that KISS1, KISS1R (GPR54), TAC3-TACR3 genes play an important role in controlling and sustaining the functioning of the reproductive system by affecting the hypothalamus-pituitary-gonad axis (HHG axis). It has been determined and investigated that it plays a role in many metabolic events such as its regulatory role in the reproductive system, its relationship with puberty and GnRH release. It has been emphasized that loss of function in existing genes, mutations may cause hypogonadism and puberty. The findings from the decidous teeth of the stem cells that carry character of, mesenchymal stem cell, associated passage number KISS1, KISS1R, TAC3, TACR3 genes to express stem cells transplanted into damaged tissue in diseases of the reproductive system are suggested as an alternative source to use it for regeneration.