Affinity-Bead-Mediated Enrichment of CD8+ Lymphocytes from Peripheral Blood Progenitor Cell Products Using Acoustophoresis

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.

Label-free separation of leukocyte subpopulations using high throughput multiplex acoustophoresis

“…Furthermore, acoustophoresis has been shown to be a gentle method which does not impact cell viability or functional capacity of the separated sample. 21,23,[27][28][29][30][31][32][33] There is a large size overlap between the different white blood cell populations with a median diameter (range) of 7.2 μm (5.5-10 μm) for lymphocytes, 9.5 μm (7.5-12 μm) for monocytes and 9.5 μm (8.5-11 μm) for granulocytes as determined by coulter counter measurements (Fig. S2 †).…”

Rapid and effective enrichment of mononuclear cells from blood using acoustophoresis

“…Size based separation was successfully shown for a variety of clinical relevant applications such as separation of lymphocytes from granulocytes 36 , isolation of tumor cells 37 , separation of WBCs from platelets 38 , cell cycle phase synchronization in mammalian cells 39 and isolation of bacteria in blood from sepsis patients 10 . Furthermore, acoustophoresis has been shown to be a gentle method that does not affect the viability and proliferative capacity of acoustically-separated cells 38 , 40 – 43 .…”

Biological Particle Control and Separation using Active Forces in Microfluidic Environments

“…They study levitation of zebrafish embryos using acoustic radiation force in a noncontact wall-less platform for a period of less than 2000 s, while the embryos still in development at 2-14 hours postfertilization, and they found that the levitation does not interfere the development, though it might influence mortality rate. Urbansky et al [40] perform the manipulation of peripheral blood progenitor cells (PBPCs) with the focus on sorting out CD8 lymphocytes (target cells) from the mixture that contains CD4, CD19, CD34, and CD56 lymphocytes as well. They label the CD8 with affinity beads, forming bead-cell complex, to modify the acoustic mobility of the target cells.…”