Large-scale isolation of CD34+ cells using the Amgen cell selection device results in high levels of purity and recovery.
The Amgen Cell Selection Device (ACSD) is a fully automated system based on the research scale magnetic-activated cell separation (MACS) system (Miltenyi Biotech GmbH, Bergisch Gladbach, Germany) for the selection of CD34+ cells. Leukapheresis products (LP) (n = 30) from normal donors mobilized with recombinant human granulocyte colony-stimulating factor (rhG-CSF) were selected with the ACSD to evaluate the performance of this system. The starting LP contained a median of 0.51% CD34+ cells (range 0.21%-1.54%) and a median WBC count of 3.0 x 10(10) (range 1-4.7 x 10(10) cells). After selection on the ACSD a mean purity of 91.5% +/- 0.6% CD34+ cells was obtained, with a median purity of 95.5% CD34+ cells. A median of 98 x 10(6) total CD34+ cells were recovered postselection, with a range of 31-323 x 10(6) cells collected from the LP. This represented a mean recovery of 81.7% +/- 6% of CD34+ cells and a median of 78% compared with starting CD34+ cell numbers in the LP. FACS analysis of the selected products demonstrated a 4-5 log depletion of T cell subsets, including CD3, CD4, CD8, and CD56 subsets. These data demonstrate the high performance obtained with the ACSD resulting in a final product of greater than 90% purity of CD34+ cells. CD34+ cells selected with the ACSD represent an ideal product for clinical applications, such as tumor cell purging, T cell depletion for allogeneic transplant, ex vivo expansion, and gene therapy.
McNiece
I
I
Department of Developmental Hematology, Amgen Inc., Thousand Oaks, California, USA.
Briddell
R
R
Stoney
G
G
Kern
B
B
Zilm
K
K
Recktenwald
D
D
Miltenyi
S
S
D015703:Antigens, CD; D018952:Antigens, CD34; D015415:Biomarkers; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D004867:Equipment Design; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000008:administration & dosage / Q000494:pharmacology*; D006410:Hematopoiesis / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007937:Leukapheresis / Q000295:instrumentation / Q000379:methods; D008212:Lymphocyte Depletion; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000494:pharmacology; D013601:T-Lymphocytes
D015703:Antigens, CD; D018952:Antigens, CD34; D015415:Biomarkers; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor