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

Background aims. Immunomagnetic enrichment of CD34+ hematopoietic "stem" cells (HSCs) using paramagnetic nano-bead 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 x 10E6 CD34+ cells, which we consider a per-kilogram dose of HSCs, products contained 17 +/- 3 x 10E3 T cells and 78 +/- 22 x 10E3 B cells. Conclusions. The process for CD34 selection with Prodigy is robust and labor-saving but not timesaving. 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.