Automated CD34+ cell isolation of peripheral blood stem cell apheresis product

  • Background aims: Immunomagnetic enrichment of CD34+ hematopoietic “stem” cells (HSCs) using paramagnetic nanobead coupled CD34 antibody and immunomagnetic extraction with the CliniMACS plus system is the standard approach to generating T-cell-depleted stem cell grafts. Their clinical beneficence in selected indications is established. Even though CD34+ selected grafts are typically given in the context of a severely immunosuppressive conditioning with anti-thymocyte globulin or similar, the degree of T-cell depletion appears to affect clinical outcomes and thus in addition to CD34 cell recovery, the degree of T-cell depletion critically describes process quality. An automatic immunomagnetic cell processing system, CliniMACS Prodigy, including a protocol for fully automatic CD34+ cell selection from apheresis products, was recently developed. We performed a formal process validation to support submission of the protocol for CE release, a prerequisite for clinical use of Prodigy CD34+ products. Methods: Granulocyte-colony stimulating factor–mobilized healthy-donor apheresis products were subjected to CD34+ cell selection using Prodigy with clinical reagents and consumables and advanced beta versions of the CD34 selection software. Target and non-target cells were enumerated using sensitive flow cytometry platforms. Results: Nine successful clinical-scale CD34+ cell selections were performed. Beyond setup, no operator intervention was required. Prodigy recovered 74 ± 13% of target cells with a viability of 99.9 ± 0.05%. Per 5 × 10E6 CD34+ cells, which we consider a per-kilogram dose of HSCs, products contained 17 ± 3 × 10E3 T cells and 78 ± 22 × 10E3 B cells. Conclusions: The process for CD34 selection with Prodigy is robust and labor-saving but not time-saving. Compared with clinical CD34+ selected products concurrently generated with the predecessor technology, product properties, importantly including CD34+ cell recovery and T-cell contents, were not significantly different. The automatic system is suitable for routine clinical application.

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Author:Gabriele Spohn, Eliza Wiercinska, Darja Karpova, Milica Bunos, Christiane Hümmer, Eva Wingenfeld, Nadine Sorg, Carolin Poppe, Volker Huppert, Juliane Stuth, Kristina Reck, Mike Essl, Erhard SeifriedORCiD, Halvard-Björn BönigORCiDGND
Pubmed Id:
Parent Title (English):Cytotherapy
Publisher:Taylor & Francis Group
Place of publication:Abington
Document Type:Article
Year of Completion:2015
Date of first Publication:2015/05/14
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2017/02/06
Tag:CD34; CliniMACS; allogeneic; automation; cell therapy; clean room; good manufacturing practice; haplo-identical; immunomagnetic; naked haplo; stem cell transplantation
Page Number:7
First Page:1465
Last Page:1471
Institutes:Medizin / Medizin
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Licence (German):License LogoCreative Commons - Namensnennung-Nicht kommerziell - Keine Bearbeitung 4.0