Abstract
Acoustophoresis is a technique that applies ultrasonic standing wave forces in a microchannel to sort cells depending on their physical properties in relation to the surrounding media. Cell handling and separation for research and clinical applications aims to efficiently separate specific cell populations. Here, we investigated the sorting of CD8 lymphocytes from peripheral blood progenitor cell (PBPC) products by affinity-bead-mediated acoustophoresis. PBPC samples were obtained from healthy donors (n = 4) and patients (n = 18). Mononuclear cells were labeled with anti-CD8-coated magnetic beads and sorted on an acoustophoretic microfluidic device and by standard magnetic cell sorting as a reference method. CD8 lymphocytes were acoustically sorted with a mean purity of 91% ± 8% and a median separation efficiency of 63% (range 15.1%–90.5%) as compared to magnetic sorting (purity 91% ± 14%, recovery 29% (range 5.1%–47.3%)). The viability as well as the proliferation capacity of sorted lymphocytes in the target fraction were unimpaired and, furthermore, hematopoietic progenitor cell assay revealed a preserved clonogenic capacity post-sorting. Bead-mediated acoustophoresis can, therefore, be utilized to efficiently sort less frequent CD8+ lymphocytes from PBPC products in a continuous flow mode while maintaining cell viability and functional capacity of both target and non-target fractions.
Highlights
Hematopoietic stem cell transplantation (HSCT) is an established therapy for hematological malignancies and other diseases
Following collection of chemotherapy- and/or hematopoietic growth factor–mobilized peripheral blood stem cells by large-volume leukapheresis [9], further processing of peripheral blood progenitor cell (PBPC) products is nowadays usually performed by large-scale magnetic cell sorting (MACS) [10,11]
We investigated the performance of affinity-bead-mediated acoustophoresis to separate CD8+ cytotoxic T cells from PBPC products and optimized the sorting for this cell population, which is less frequent in the starting material compared to CD4+ cells
Summary
Hematopoietic stem cell transplantation (HSCT) is an established therapy for hematological malignancies and other diseases. Label-free separation of lymphocytes and granulocytes as well as platelets was demonstrated using microchip-based free-flow acoustophoresis [25,27] Based on these results we developed a system to acoustically sort bead-labeled CD4 lymphocytes from peripheral stem cell products [28]. Affinity-bead-mediated acoustophoresis can be used to target and enrich specific cell populations in a continuously perfused microfluidic device These data represent a step forward towards acoustic sorting of even rarer cell populations, such as peripheral blood stem cells and open up the potential for simultaneous acoustic separation of multiple cell populations
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