Detecting primitive hematopoietic stem cells in total nucleated and mononuclear cell fractions from umbilical cord blood segments and units

Journal of Translational Medicine
John PattersonIvan N Rich

Abstract

Rare hematopoietic stem cell populations are responsible for the transplantation engraftment process. Umbilical cord blood (UCB) is usually processed to the total nucleated cell (TNC), but not to the mononuclear cell (MNC) fraction. TNC counts are used to determine UCB unit storage, release for transplantation and correlation with time to engraftment. However, the TNC fraction contains varying concentrations of red blood cells, granulocytes, platelets and other cells that dilute and mask the stem cells from being detected. This does not allow the quality and potency of the stem cells to be reliably measured. 63 UCB segments and 10 UCB units plus segments were analyzed for the response of both primitive lympho-hematopoietic and primitive hematopoietic stem cells in both the TNC and MNC fractions. The samples were analyzed using a highly sensitive, standardized and validated adenosine triphosphate (ATP) bioluminescence stem cell proliferation assay verified against the colony-forming unit (CFU) assay. Dye exclusion and metabolic viability were also determined. Regardless of whether the cells were derived from a segment or unit, the TNC fraction always produced a significantly lower and more variable stem cell response than that d...Continue Reading

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Citations

Mar 13, 2016·Revista brasileira de hematologia e hemoterapia·José Carlos Jaime-PérezAlejandra Celina Esparza-Sandoval
Apr 5, 2016·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Santiago RouraAntoni Bayes-Genis
Jul 15, 2015·Stem Cells Translational Medicine·Ivan N Rich
Nov 28, 2018·Journal of Cellular Physiology·Behnaz TaheriNosratollah Zarghami

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Methods Mentioned

BETA
density gradient centrifugation
flow cytometry
Assay

Software Mentioned

GraphPad Prism
Systat

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