| Popis: |
Objectives: Hematopoietic stem cell transplantation (HSCT) is a widely used method for treatment of hematological disorders and some other diseases. However, sometimes a suitable donor of hematopoietic stem cells (HSC) is not found for a patient. Because HSC have been described as cells with low proliferative and metabolic activity, their tolerance to the lack of oxygen or metabolic substrates may be assumed. In this study, we explored cadaveric bone marrow as an alternative source of HSC for HSCT, using a mouse experimental model. In addition, the effect of in vitro metabolic inhibition and short-term in vitro storage (1 - 4 days) on functional properties of mouse HSC was investigated. Methods: C57Bl/6 mice (wild-type or p53-/- ) were used in the experiments. To explore cadaveric HSC, bone marrow (BM) was left in intact femurs at 37řC, 20řC and 4řC under the conditions of ischemia. The bone marrow cells were harvested after defined time periods ranging 0 - 48 hours. For metabolic inhibition, the electron transport chain inhibitor potassium cyanide (KCN) and inhibitor of glycolysis 2-deoxy-D-glucose (2-DG) were used in vitro. To determine the impact of ischemia, metabolic inhibition, or in vitro storage on transplantability of HSC, the competitive repopulation assay using Ly5.1/Ly5.2 congenic model... |
