Quality testing of an innovative cascade separation system for multiple cell separation

Arkadiusz Pierzchalski; Aleksandra Moszczynska; Bernd Albrecht; Jan-Michael Heinrich; Attila Tarnok

doi:10.1117/12.906412

9 February 2012 Quality testing of an innovative cascade separation system for multiple cell separation

Arkadiusz Pierzchalski, Aleksandra Moszczynska, Bernd Albrecht, Jan-Michael Heinrich, Attila Tarnok

Proceedings Volume 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X; 82250O (2012) https://doi.org/10.1117/12.906412
Event: SPIE BiOS, 2012, San Francisco, California, United States

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.

Citation Download Citation

Arkadiusz Pierzchalski, Aleksandra Moszczynska, Bernd Albrecht, Jan-Michael Heinrich, and Attila Tarnok "Quality testing of an innovative cascade separation system for multiple cell separation", Proc. SPIE 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X, 82250O (9 February 2012); https://doi.org/10.1117/12.906412

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
12 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Blood

Statistical analysis

Magnetism

Software development

Contamination

Data analysis

Manufacturing

Show All Keywords

Keywords/Phrases

Search In:

Publication Years