Isolation of monocytes from whole blood-derived buffy coats by continuous counter-flow elutriation

• Published in: Journal of clinical apheresis 2006-10, Vol.21 (3), p.153-157
• Main Authors: Schwanke, Uwe, Nabereit, Anja, Moog, Rainer
• Format: Article
• Language: English
• Subjects: Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Biological and medical sciences; Blood Component Removal - instrumentation; Blood Component Removal - methods; Blood. Blood and plasma substitutes. Blood products. Blood cells. Blood typing. Plasmapheresis. Apheresis; Bone marrow, stem cells transplantation. Graft versus host reaction; buffy coat; Cell Separation - methods; Cobe Spectra; continuous counter-flow elutriation; Dendritic Cells - cytology; Elutra; Emergency and intensive care: renal failure. Dialysis management; Humans; Intensive care medicine; Medical sciences; monocytes; Monocytes - cytology; Neutrophils - cytology; Time Factors; Transfusions. Complications. Transfusion reactions. Cell and gene therapy
• ISSN: 0733-2459; 1098-1101
• DOI: 10.1002/jca.20077

Monocytes (MOs) are the most commonly used precursors for the generation of dendritic cells (DCs) in vitro. Continuous counter‐flow elutriation represents a promising tool to isolate MOs from white blood cell (WBC) products. Thirty whole blood–derived, AB0‐identical buffy coats (BCs) were pooled using sterile technique (n = 5 experiments). For red blood cell (RBC) and polymorphonuclear cell (PMN) depletion, the BC pools were processed in a Cobe® Spectra device (Gambro BCT) using the bone marrow program. Subsequently, continuous counter‐flow elutriation in an Elutra® device (Gambro BCT) was performed to enrich and purify MOs. BC pool volume averaged 1,260 ± 14 ml containing 7.7 ± 1.1 × 109 MOs. During 107 ± 7 min, Cobe Spectra operation, the BC pools were processed for several times, and approximately 9,749 ± 605 ml volume passed the device. Product volume and MO yield averaged 160 ± 16 ml, and 4.3 ± 1.3 × 109 cells, respectively. Elutra operation was performed within 59 ± 0 min and yielded 2.5 ± 0.9 × 109 MOs with a purity of 60 ± 12%. Compared with the Cobe Spectra product cell count, MO recovery by Elutra averaged 59 ± 10%. Elutriation of MOs from pooled BCs using Elutra exhibited comparatively low recovery and purity rates. This shortcoming may be due to the nature of the source material. Optimization of the elutriation procedure is necessary to improve MO enrichment from BCs. J. Clin. Apheresis 2006. © 2006 Wiley‐Liss, Inc.