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Differentiation Kinetics of Chronic Hypoxia Hematopoietic Stem Cells

In vivo, stem cells can be found in hypoxic niches within bone marrow, peripheral blood, and umbilical cord blood. Stem cells grown in hypoxic conditions, which contains an oxygen level of up to 1%, cause them to utilize glycolysis as the primary source of energy, as opposed to stem cells cultured in normoxic conditions that use both glycolysis and oxidative phosphorylation. Differentiation requires a significant amount of energy, and previous research has shown that hydroxide radicals leaked from mitochondria inhibit phosphatases and stimulate differentiation. EML cells are a stem cell factor dependent murine hematopoietic stem cell line often used to study differentiation of blood cells. In our study, EML cells were allowed to differentiate in normoxia, chronic hypoxia and acute hypoxia conditions. We observed the effects that each of the conditions have on the differentiation kinetics by using antibodies to label the cell surface markers against: Sca‐1, CD117, CD48, CD135, CD150, CD11b, CD127, F4/80, Ter119, Ly6‐G/C. Chronic hypoxia EML cells differentiation kinetics were significantly altered. Also, labeling cells with fluorescent antibodies allowed us to trace the lineage of the induced differentiated cells, by using flow cytometry. We found that EML cells cultured in chronic hypoxic conditions had a shift in its multi‐potential progenitor (MPP) subtype, MPP4 to MPP3, that the other conditions did not. Further investing the differentiation kinetics of chronic hypoxia could lead to a better understanding on producing long term hematopoietic stem cells in vitro, for stem cell transplantation in healthcare.