True volumetric method for flow cytometric enumeration of CD34 + stem cells and its agreement with a standard bead-based single-platform protocol

Cytotherapy. 2012 May;14(5):621-9. doi: 10.3109/14653249.2012.667875. Epub 2012 Mar 20.

Abstract

Background aims: Stem cells are commonly enumerated with bead-based methods in blood and marrow progenitor cell transplantation centers. We compared the International Society of Hematotherapy and Graft Engineering (ISHAGE) bead-based method with a true volumetric one that obviates the use of fluorescent beads for enumeration.

Methods: From 31 samples, including 15 peripheral blood samples and 16 leukapheresis products, CD34 (+) cells were enumerated with the single-platform bead-based ISHAGE method and a true volumetric method. After exclusion of two outliers, one from the peripheral blood group and the other from the leukapheresis group, the results were compared.

Results: In the peripheral blood category, no significant difference was observed. However, a proportional systematic error was seen in the leukapheresis group. The systematic error was corrected in the leukapheresis group using a regression line equation. The 95% confidence interval of differences was [-5.83, 2.18] for the peripheral blood and [-38.40, 38.77] for the leukapheresis group after correction of the systematic error.

Conclusions: The true volumetric method is a simple and reliable approach that can be used instead of the more popular bead-based procedures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antigens, CD34 / blood*
  • Bone Marrow Transplantation*
  • Female
  • Flow Cytometry*
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / metabolism
  • Hodgkin Disease / blood*
  • Hodgkin Disease / therapy
  • Humans
  • Leukapheresis
  • Leukocyte Count
  • Lymphoma, Non-Hodgkin / blood*
  • Lymphoma, Non-Hodgkin / therapy
  • Male
  • Middle Aged
  • Multiple Myeloma / blood*
  • Multiple Myeloma / therapy

Substances

  • Antigens, CD34