Rapid debulking and CD34 enrichment of filgrastim-mobilized peripheral blood stem cells by semiautomated density gradient centrifugation in a closed system.
Filgrastim-mobilized peripheral blood progenitor cells (PBPC) are used for hematopoietic reconstitution after myeloablative therapy for malignancies, but the large number of cells collected in a single apheresis frequently presents a problem for storage or further processing. We have evaluated the use of CD34 Buoyant Density Solution-PBPC, an ultralight-density colloidal silica suspension, for debulking and enrichment of CD34+ cells in PBPC preparations in a semiautomated system. Cells were collected from four filgrastim-treated normal donors using the COBE Spectra. The separation procedure was carried out with Plasma-Lyte A and DNase 5 U/ml using the COBE 2991. Following processing and washing, there was a 26% recovery of nucleated cells, 2.6-fold enrichment of CD34+ cells, 68% recovery of CD34+ cells, 88% recovery of CFU-GM, 73% recovery of BFU-E, 1 log depletion of CD3+ cells, 0.5 log depletion of CD56+ cells, and 1 log depletion of CD19+ cells. These results were not significantly different from those obtained when PBPC were separated over CD34 Buoyant Density Solution-PBPC by centrifugation in tubes. Using CD34 Buoyant Density Solution-PBPC, mononuclear preparations of PBPC can be enriched rapidly for CD34+ cells and depleted of lymphocytes in a semiautomated closed system using reagents produced according to good manufacturing practice (GMP).
Przepiorka
D
D
University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Van Vlasselaer
P
P
Huynh
L
L
Durett
A
A
Agbor
P
P
Lauppe
J
J
Valone
F
F
Champlin
R
R
Körbling
M
M
D018952:Antigens, CD34; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D003470:Culture Media; D000069585:Filgrastim; D016179:Granulocyte Colony-Stimulating Factor; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D011994:Recombinant Proteins
D018952:Antigens, CD34; D003470:Culture Media; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D000069585:Filgrastim