title: Automated CD34+ cell isolation of peripheral blood stem cell apheresis product.
abstract: OBJECTIVE
  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.
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Department of Cellular Therapeutics, Frankfurt, Germany.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Spohn
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Department of Cellular Therapeutics, Frankfurt, Germany.
authors.forename: Eliza
authors.initials: E
authors.lastname: Wiercinska
authors.affiliation: Goethe University Medical Center, Institute for Transfusion Medicine and Immunohematology, Frankfurt, Germany.
authors.forename: Darja
authors.initials: D
authors.lastname: Karpova
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Department of Cellular Therapeutics, Frankfurt, Germany.
authors.forename: Milica
authors.initials: M
authors.lastname: Bunos
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Department of Cellular Therapeutics, Frankfurt, Germany.
authors.forename: Christiane
authors.initials: C
authors.lastname: Hümmer
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Department of Cellular Therapeutics, Frankfurt, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Wingenfeld
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Department of Cellular Therapeutics, Frankfurt, Germany.
authors.forename: Nadine
authors.initials: N
authors.lastname: Sorg
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Department of Cellular Therapeutics, Frankfurt, Germany.
authors.forename: Carolin
authors.initials: C
authors.lastname: Poppe
authors.affiliation: Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Huppert
authors.affiliation: Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Stuth
authors.affiliation: Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany.
authors.forename: Kristina
authors.initials: K
authors.lastname: Reck
authors.affiliation: Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany.
authors.forename: Mike
authors.initials: M
authors.lastname: Essl
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Department of Cellular Therapeutics, Frankfurt, Germany; Goethe University Medical Center, Institute for Transfusion Medicine and Immunohematology, Frankfurt, Germany.
authors.forename: Erhard
authors.initials: E
authors.lastname: Seifried
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Institute Frankfurt, Department of Cellular Therapeutics, Frankfurt, Germany; Goethe University Medical Center, Institute for Transfusion Medicine and Immunohematology, Frankfurt, Germany; University of Washington, Department of Medicine, Division of Hematology, Seattle, Washington, USA. Electronic address: h.boenig@blutspende.de.
authors.forename: Halvard
authors.initials: H
authors.lastname: Bönig
mesh_terms: D018952:Antigens, CD34 / Q000276:immunology*; D000961:Antilymphocyte Serum / Q000276:immunology; D057205:Automation, Laboratory; D001402:B-Lymphocytes / Q000276:immunology; D001781:Blood Component Removal / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D008212:Lymphocyte Depletion / Q000379:methods; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D018952:Antigens, CD34; D000961:Antilymphocyte Serum; D016179:Granulocyte Colony-Stimulating Factor
keywords: CD34; CliniMACS; Prodigy; allogeneic; automation; cell therapy; clean room; good manufacturing practice; haplo-identical; immunomagnetic; naked haplo; stem cell transplantation