Preliminary evaluation of a highly automated instrument for the selection of CD34+ cells from mobilized peripheral blood stem cell concentrates.
BACKGROUND
Cell selection is an important part of manufacturing cellular therapies. A new highly automated instrument, the CliniMACS Prodigy (Miltenyi Biotec), was evaluated for the selection of CD34+ cells from mobilized peripheral blood stem cell (PBSC) concentrates using monoclonal antibodies conjugated to paramagnetic particles.


METHODS
PBSCs were collected by apheresis from 36 healthy subjects given granulocyte-colony-stimulating factor (G-CSF) or G-CSF plus plerixafor. CD34+ cells from 11 PBSC concentrates were isolated with the automated CliniMACS Prodigy and 25 with the semiautomated CliniMACS Plus Instrument.


RESULTS
The proportion of CD34+ cells in the selected products obtained with the two instruments was similar: 93.6 ± 2.6% for the automated and 95.7 ± 3.3% for the semiautomated instrument (p > 0.05). The recovery of CD34+ cells from PBSC concentrates was less for the automated than the semiautomated instrument (51.4 ± 8.2% vs. 65.1 ± 15.7%; p = 0.019). The selected products from both instruments contained few and similar quantities of platelets (PLTs) and red blood cells. The depletion of CD3+ cells was less with the automated instrument (4.34 ± 0.2 log depletion vs. 5.20 ± 0.35 log depletion; p < 1 × 10(-6) ). Removal of PLTs from PBSC concentrates by washing was associated with better CD34+ cell recovery. We explored the reasons for lower CD34+ cell recovery by the Prodigy and found that the nonselected cells for the Prodigy contained more PLTs than those for the CliniMACS Plus.


CONCLUSIONS
CD34+ cells can be effectively selected from mobilized PBSC concentrates with the CliniMAC Prodigy, but the recovery of CD34+ cells and depletion of CD3+ cells was lower than with the semiautomated CliniMACS Plus Instrument.
Cell Processing Section, Department of Transfusion Medicine, Clinical Center.
David F
DF
Stroncek
Cell Processing Section, Department of Transfusion Medicine, Clinical Center.
Minh
M
Tran
Cell Processing Section, Department of Transfusion Medicine, Clinical Center.
Sue Ellen
SE
Frodigh
Cell Processing Section, Department of Transfusion Medicine, Clinical Center.
Virginia
V
David-Ocampo
Cell Processing Section, Department of Transfusion Medicine, Clinical Center.
Jiaqiang
J
Ren
National Heart, Lung and Blood Institute.
Andre
A
Larochelle
Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases.
Virginia
V
Sheikh
Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases.
Irini
I
Sereti
Molecular Genomics and Therapeutics Section, Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland.
Jeffery L
JL
Miller
Miltenyi Biotec, Auburn, California.
Kevin
K
Longin
Cell Processing Section, Department of Transfusion Medicine, Clinical Center.
Marianna
M
Sabatino
D018952:Antigens, CD34 / Q000097:blood; D001596:Benzylamines; D001781:Blood Component Removal / Q000295:instrumentation* / Q000379:methods*; D000080027:Cyclams; D005260:Female; D016179:Granulocyte Colony-Stimulating Factor / Q000008:administration & dosage*; D019650:Hematopoietic Stem Cell Mobilization; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006571:Heterocyclic Compounds / Q000008:administration & dosage*; D006801:Humans; D008297:Male
D018952:Antigens, CD34; D001596:Benzylamines; D000080027:Cyclams; D006571:Heterocyclic Compounds; D016179:Granulocyte Colony-Stimulating Factor; C088327:plerixafor