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Proceedings Paper

Quality testing of an innovative cascade separation system for multiple cell separation
Author(s): Arkadiusz Pierzchalski; Aleksandra Moszczynska; Bernd Albrecht; Jan-Michael Heinrich; Attila Tarnok
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Paper Abstract

Isolation of different cell types from mixed samples in one separation step by FACS is feasible but expensive and slow. It is cheaper and faster but still challenging by magnetic separation. An innovative bead-based cascade-system (pluriSelect GmbH, Leipzig, Germany) relies on simultaneous physical separation of different cell types. It is based on antibody-mediated binding of cells to beads of different size and isolation with sieves of different mesh-size. We validated pluriSelect system for single parameter (CD3) and simultaneous separation of CD3 and CD15 cells from EDTA blood-samples. Results were compared with those obtained by MACS (Miltenyi-Biotech) magnetic separation (CD3 separation). pluriSelect separation was done in whole blood, MACS on Ficoll gradient isolated leukocytes, according to the manufacturer's protocols. Isolated and residual cells were immunophenotyped (7-color 8-antibody panel (CD3; CD16/56; CD4; CD8; CD14; CD19; CD45; HLADR) on a CyFlowML flow cytometer (Partec GmbH). Cell count (Coulter), purity, yield and viability (7-AAD exclusion) were determined. There were no significant differences between both systems regarding purity (92-98%), yield (50-60%) and viability (92-98%) of isolated cells. PluriSelect separation was slightly faster than MACS (1.15 h versus 1.5h). Moreover, no preenrichment steps were necessary. In conclusion, pluriSelect is a fast, simple and gentle system for efficient simultaneous separation of two cell subpopulation directly from whole blood and can provide a simple alternative to FACS. The isolated cells can be used for further research applications.

Paper Details

Date Published: 9 February 2012
PDF: 12 pages
Proc. SPIE 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X, 82250O (9 February 2012); doi: 10.1117/12.906412
Show Author Affiliations
Arkadiusz Pierzchalski, Univ. Leipzig (Germany)
Cardiac Ctr. GmbH (Germany)
Aleksandra Moszczynska, Univ. Leipzig (Germany)
Cardiac Ctr. GmbH (Germany)
Bernd Albrecht, Cardiac Ctr. GmbH (Germany)
Jan-Michael Heinrich, pluriSelect GmbH (Germany)
Attila Tarnok, Univ. Leipzig (Germany)
Cardiac Ctr. GmbH (Germany)


Published in SPIE Proceedings Vol. 8225:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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