Improved expansion of T cells in culture when isolated with an equipment-free, high-throughput, flow-through microfluidic module versus traditional density gradient centrifugation.

Autor: Strachan BC; Department of Biomedical Engineering, University of Houston, Houston, Texas, USA., Xia H; Department of Biomedical Engineering, University of Houston, Houston, Texas, USA., Vörös E; Department of Biomedical Engineering, University of Houston, Houston, Texas, USA., Gifford SC; Halcyon Biomedical Incorporated, Friendswood, Texas, USA., Shevkoplyas SS; Department of Biomedical Engineering, University of Houston, Houston, Texas, USA. Electronic address: sshevkoplyas@uh.edu.
Jazyk: angličtina
Zdroj: Cytotherapy [Cytotherapy] 2019 Feb; Vol. 21 (2), pp. 234-245. Date of Electronic Publication: 2019 Jan 16.
DOI: 10.1016/j.jcyt.2018.12.004
Abstrakt: Background: The isolation of lymphocytes - and removal of platelets (PLTs) and red blood cells (RBCs) - from an initial blood sample prior to culture is a key enabling step for effective manufacture of cellular therapies. Unfortunately, currently available methods suffer from various drawbacks, including low cell recovery, need for complex equipment, potential loss of sterility and/or high materials/labor cost.
Methods: A newly developed system for selectively concentrating leukocytes within precisely designed, but readily fabricated, microchannels was compared with conventional density gradient centrifugation with respect to: (i) ability to recover lymphocytes while removing PLTs/RBCs and (ii) growth rate and overall cell yield once expanded in culture.
Results: In the optimal embodiment of the new microfluidic approach, recoveries of CD3+, CD19+ and CD56+ cells (85%, 89% and 97%, respectively) were significantly higher than for paired samples processed via gradient-based separation (51%, 53% and 40%). Although the removal of residual PLTs and RBCs was lower using the new approach, its enriched T-cell fraction nevertheless grew at a significantly higher rate than the gradient-isolated cells, with approximately twice the cumulative cell yield observed after 7 days of culture.
Discussion: The standardization of each step of cellular therapy manufacturing would enable an accelerated translation of research breakthroughs into widely available clinical treatments. The high-throughput approach described in this study - requiring no ancillary pumping mechanism nor expensive disposables to operate - may be a viable candidate to standardize and streamline the initial isolation of lymphocytes for culture while also potentially shortening the time required for their expansion into a therapeutic dose.
(Copyright © 2018 International Society for Cell and Gene Therapy. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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