Purification of monocytes from cryopreserved mobilized apheresis products by elutriation with the Elutra device.
The Elutra biomedical device allows semi-automatic enrichment of monocytes by elutriation, using a single-use, closed and cGMP compliant tubing set, in a cost effective way. The procedure has been validated using fresh apheresis products from nonmobilized donors. We here evaluated the possibility of using Elutra to enrich monocytes from frozen/thawed apheresis products collected from mobilized healthy donors. Frozen apheresis products from 6 G CSF mobilized donors were thawed and used in 16 elutriation procedures. We compared the recovery and purity of enriched monocytes using different buffer compositions and elutriation profiles. Elutriated monocytes were cultured to generate mature dendritic cells (DCs). Depending in part of the initial granulocyte contamination in the apheresis product, the use of Desoxyribo Nuclease (DNAse) to avoid aggregation, was needed through only the initial steps or throughout the elutriation process. The average monocyte recovery was 85+/-31%. The average purity was 73+/-9%. The recovery of mature DC at d8 of culture was 20+/-6% of the input monocyte numbers. We conclude that Elutra allows the purification of monocytes from thawed mobilized apheresis. It requires no pre-processing of the cell product before elutriation, and allows the generation of phenotypically mature DC in quantities that are compatible with a clinical use.
Lemarie
Claude
C
The MD Anderson Cancer Center, Blood and Marrow Transplantation Department, Houston, Texas, USA. lemariec@marseille.fnclcc.fr
Sugaye
Romina
R
Kaur
Indreshpaul
I
Taga
Tim
T
Chabannon
Christian
C
Schuyler
Robert
R
Mcmannis
John
J
D015703:Antigens, CD / Q000032:analysis; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods; D015925:Cryopreservation / Q000379:methods*; D003713:Dendritic Cells / Q000737:chemistry / Q000166:cytology; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology; D007937:Leukapheresis; D009000:Monocytes / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D009504:Neutrophils / Q000166:cytology; D011994:Recombinant Proteins
D015703:Antigens, CD; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor