Purification of regulatory T cells with the use of a fully enclosed high-speed microfluidic system |
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| Affiliation: | 1. Cytonome/ST, LLC, Boston, Massachusetts, USA;2. Bauer Flow Cytometry Core Facility, FAS Center for Systems Biology, Harvard University, Cambridge, Massachusetts, USA;3. Department of Immunology, Genetics and Pathology, Section of Transplantation Surgery, Uppsala University, Uppsala, Sweden;4. Department of Surgical Sciences, Section of Transplantation Surgery, Uppsala University, Uppsala, Sweden;1. Anthony Nolan Research Institute and University College London, Royal Free Campus, London, United Kingdom;2. Haematology Department, Hammersmith Hospital, London, United Kingdom;1. Division of Plastic and Hand Surgery, University Hospital Zurich, Zurich, Switzerland;2. Department of Clinical Research, University of Bern, Bern, Switzerland;3. Center for Clinical Research, University of Zurich, Zurich, Switzerland;4. Department of Ophthalmology, University of Bern, Inselspital, Bern, Switzerland;1. Department of Chemical and Biological Engineering, Koc University, Istanbul, 34450, Turkey;2. Department of Physics, Koc University, Istanbul, 34450, Turkey;3. Department of Mechanical Engineering, Koc University, Istanbul, 34450, Turkey;4. Department of Electrical and Electronics Engineering, Koc University, Istanbul, 34450, Turkey;5. Koç University TÿPRAſ Energy Center (KUTEM), Koç University, 34450 Sarıyer, Istanbul, Turkey;1. Department of Orthopedics, Korea University Ansan Hospital, Ansan, South Korea;2. Next-Generation Pharmaceutical Research Center, Korea Institute of Toxicology, Daejeon, Republic of Korea;3. Scoliosis Research Institute, Department of Orthopedics, Korea University Guro Hospital, Seoul, South Korea;1. Oncohematology Research Group, Navarrabiomed, Miguel Servet Foundation, Pamplona, Spain;2. Department of Haematology, Complejo Hospitalario de Navarra, Navarra Health Service, Pamplona, Spain;3. Department of Haematology, University College London Medical School, University College London, London, United Kingdom |
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| Abstract: | Background aimsDespite promising advances in cellular therapies, it will be difficult to fully test or implement new therapies until advances are made in the processes for cell preparation. This study describes the use of an advanced prototype of a flow-cytometry cell purification system constructed for operation in a clinical environment to prepare regulatory T cells defined as CD4+/CD25bright/CD127neg/low.MethodsThe sort performance of the Gigasort system was directly compared with available droplet sorters using mixtures of highly fluorescent and non-fluorescent 5-μm polystyrene particles. CD4+-enriched cell preparations were processed with the use of a sterile, disposable fluid handling unit with a chip containing parallel microfluidic-based sorters.ResultsSimilar purity and sort efficiency as found with droplet sorters were obtained with the 24-channel chip sorter system. Starting with 450 million fresh peripheral blood mononuclear cells, 150,000 to 1.7 million cells that were, on average, 85% FoxP3-positive and 97% viable, were obtained in <4 h.ConclusionsThis study presents a technology adapted to regulatory requirements for clinical cell purification and that achieves high throughput and cell-friendly conditions by use of a microfluidic chip with 24 parallel microsorters, providing a rapid, sterile method of purifying regulatory T cells accurately and with excellent viability. |
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| Keywords: | cell purification cell separation immunotherapy regulatory T cells |
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