Elimination of cytotoxic and genotoxic barriers during course of HSC expansion with pifithrin alpha treatment

Hematopoietic stem cells (HSCs) are used in the treatment of a extensive range of blood diseases. However, the limited number and stagnation feature make it difficult to use in stem cell treatment. Numerous studies have been carried out with small molecules to ensure the expansion and multiplication of HSCs ex vivo. As known; Genetic inhibition of P53 has been shown to lead to the proliferation of hematopoietic stem cells (HSCs). The pifithrin-apla is a small molecule compound and the inhibitor of p53 function and is known to be a precaution against genotoxic agents. In this study; The effects of the pifithrin-apla molecule on HSC cells were investigated. For this purpose, mouse HSCs were enriched by magnetic line abstraction. Human umbilical cord blood (UCB) and osseous (BM) mononuclear cells were isolated by histopak intensity gradient centrifugation. When the cells were cultured under appropriate conditions with the prepared HSC media, DMSO treatment was used as a control. After seven days of incubation with the small molecule of pifithrin-apla, mouse HSCs were analyzed for lin (-), sca1, c-kit, MD34 markers. Likewise, for human HSCs, CD34 and CD133 markers were analyzed. Cell line analysis and apoptosis analysis were performed following incubation of FACS-purified mouse LSK cells with the selected optimal dose molecule. At the same time, the effects of Human Umbilical Venous Endothelial Cells (HUVEC) and Human Dermal Fibroblast (HDF) and fat tissue on MSCs were also examined and did not have a negative effect. The small molecule of pifithrin-alpha caused a 3-fold increase in LSK (lin-sca1 + c-kit +) and human UCB HSC cells. Similarly, human BM caused a 2-fold increase in HSC cells. At the same time, chicken HSC was isolated and surface marker secreening was performed. So; Avian influenza and the effects of human HSCs. As a result, it is thought that this molecule can be used to increase ex vivo HSC expansion and treatment efficacy.