Debulking blood stem cell collections by density gradient centrifugation in a closed-vessel system

Cytotherapy. 1999;1(2):111-7. doi: 10.1080/0032472031000141248.


Background: Blood stem cells collected by apheresis are largely mononuclear cells in nature, so manipulation of blood stem-cell components frequently requires more time and reagents than for a marrow harvest. Reducing the nucleated cell number in mobilized blood stem-cell collections, while preserving hematopoietic progenitor content, would make such manipulations simpler and less costly, but only if debulking procedures were not complex.

Methods: We evaluated separation of light-density cells and enrichment of CD34+ cells from mobilized peripheral blood stem-cell collections by density gradient centrifugation over buoyant density solution 60 (BDS 60) in a single, closed vessel.

Results: Fifteen apheresis products from five normal volunteers and eight cancer patients contained 3.44 (range, 1.19-5.51) x 10(10) nucleated cells. Following processing and washing, there was a median 29% recovery of nucleated cells, 79% recovery of CD34+ cells, 2.49-fold enrichment of CD34+ cells, 0.96-log depletion of CD3+ cells, 0.48-log depletion of CD56+ cells, and 0.72-log depletion of CD19+ cells. Results of processing were affected by the variability in composition of the apheresis products. The enrichment of CD34+ cells varied by donor type, and there was a logarithmic relationship between the preprocessing percentage of CD19+ cells and the log reduction in CD19+ cells. Recovery of cells after thawing and washing was acceptable for processed cells cryopreserved at concentrations over the range of 0.01-1.5 x 10(8)/mL.

Discussion: These results demonstrate a simple method by which an apheresis product of 1-5 x 10(10) cells can be debulked effectively in a single, closed vessel.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Antigens, CD19 / analysis
  • Antigens, CD34 / analysis
  • Blood Cell Count
  • Blood Component Removal*
  • CD3 Complex / analysis
  • CD56 Antigen / analysis
  • Cell Separation / instrumentation
  • Cell Separation / methods*
  • Centrifugation, Density Gradient* / instrumentation
  • Cryopreservation
  • Equipment Design
  • Female
  • Filgrastim
  • Granulocyte Colony-Stimulating Factor / therapeutic use
  • Hematopoietic Stem Cell Mobilization
  • Hematopoietic Stem Cell Transplantation / methods*
  • Hematopoietic Stem Cells / drug effects
  • Hematopoietic Stem Cells / immunology
  • Hematopoietic Stem Cells / physiology*
  • Humans
  • Male
  • Middle Aged
  • Recombinant Proteins
  • Young Adult


  • Antigens, CD19
  • Antigens, CD34
  • CD3 Complex
  • CD56 Antigen
  • Recombinant Proteins
  • Granulocyte Colony-Stimulating Factor
  • Filgrastim