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: | Hui Xia, Eszter Vörös, Briony C. Strachan, Sean C. Gifford, Sergey S. Shevkoplyas |
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| Rok vydání: | 2019 |
| Předmět: |
Blood Platelets
0301 basic medicine Cancer Research Erythrocytes Density gradient Cell Survival T-Lymphocytes Microfluidics Immunology Cell Count Cell Separation Article CD19 03 medical and health sciences 0302 clinical medicine Paired samples Centrifugation Density Gradient Humans Immunology and Allergy Throughput (business) Cell yield Cells Cultured Genetics (clinical) Differential centrifugation Transplantation biology Chemistry Cell Biology Leukapheresis Adoptive Transfer 030104 developmental biology Oncology 030220 oncology & carcinogenesis biology.protein Filtration Biomedical engineering |
| Zdroj: | Cytotherapy. 21:234-245 |
| ISSN: | 1465-3249 |
| DOI: | 10.1016/j.jcyt.2018.12.004 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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