Measurement of mitochondrial activity in apheresis collection of Hematopoietic stem cells samples

Hematopoietic stem cells (HSCs) develop into all types of blood cells. HSCs rely on glycolysis for energy production, HSCs that do not differentiate hardly use mitochondrial oxidative phosphorylation (OxPHOS) - therefore they have a relatively low metabolic activity but during the differentiation of HSCs, a rapid transition from glycolysis to the production of OxPHOS and mitochondrial ATP occurs and a relatively high metabolic activity is created. Transplantation of autologous stem cells forms the basis of cancer treatment. In the paresis method, the stem cells are mobilized into the peripheral blood with mobilization. During collection, the machine collects all blood components and separates them into low-density mononuclear cells. At the end of the collection a CD34 count is taken and checked whether the collection meets the standard of 1.5 cells/kg. However, the mitochondrial activity of the cells was not tested.
Samples were obtained from apheresis product of 27 patients with blood cancer undergoing autologous transplantation who signed an informed consent form. The cells were stained with Antibodies CD34-Fitc, CD38-PEcy7, TMRE-PE, DAPI-PB. FACS cytoflex is used for analysis and Kaluza Analysis software was used to analyze FACS data.
Our results showed 3 population of DAPI negative that analyzed to TMRE.
( CD38+CD34-, CD38+CD34+ CD38-CD34+) The population of CD38+CD34- showed the high florasent intensity. Our data observed that different populations have different membrane potentials and there is a variation from patient to patient.