The use of acoustic force capture to ultra-purify lymphocyte subpopulations from human adult whole blood.

Abstract

Pre-analytical enrichment of cell populations prior to therapeutic delivery is of paramount interest throughout the fields of regenerative medicine and clinical interventional therapies. Enrichment of a cell population typically involves two aspects: (a) the increase in concentration of particular subpopulation of the total cell fraction by means of removal of other cells of no interest to the particular interrogation; and (b) improvement of resolution of signal by removal of 'noise' mostly arising from cellular debris in the treated sample. In this research, leukocyte populations were obtained from erythrocyte-depleted primary whole blood from human adults and subjected to flow through acoustic fields within the ultrasound range to remove cellular debris. It was possible to demonstrate aggregation and holding of leukocytes by using ultrasound within the frequency range 11.448-11.483 MHz, which facilitated removal of cellular debris by washing under continuous perfusion. The T-lymphocyte population were phenotypically characterized using CD4/CD8 (T(h)/T(c)) immunocytochemistry by flow cytometry and demonstrated a significant decrease in 'false-positive' events during cellular analysis, due to the efficient eradication of non-specifically reactive cells and tissue debris from the cell populations of interest. Therefore, it was possible to conclude that flow through an ultrasonic acoustic system was capable of providing a non-destructive method for the hyper-purification of primary derived cell populations, with potential exploitations throughout the fields of cellular research, medical diagnostics and clinical therapies.