title: Purification of regulatory T cells with the use of a fully enclosed high-speed microfluidic system.
abstract: OBJECTIVE
  Despite 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(+)/CD25(bright)/CD127(neg/low).
  
  
  METHODS
  The 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.
  
  
  RESULTS
  Similar 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.
  
  
  CONCLUSIONS
  This 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.
authors: Cytonome/ST, LLC, Boston, Massachusetts, USA. Electronic address: rhulspas@cytonome.com.
authors: Ruud
authors: R
authors: Hulspas
authors: Cytonome/ST, LLC, Boston, Massachusetts, USA.
authors: Lydia
authors: L
authors: Villa-Komaroff
authors: Cytonome/ST, LLC, Boston, Massachusetts, USA.
authors: Erin
authors: E
authors: Koksal
authors: Cytonome/ST, LLC, Boston, Massachusetts, USA.
authors: Kenol
authors: K
authors: Etienne
authors: Bauer Flow Cytometry Core Facility, FAS Center for Systems Biology, Harvard University, Cambridge, Massachusetts, USA.
authors: Patricia
authors: P
authors: Rogers
authors: Cytonome/ST, LLC, Boston, Massachusetts, USA.
authors: Matt
authors: M
authors: Tuttle
authors: Department of Immunology, Genetics and Pathology, Section of Transplantation Surgery, Uppsala University, Uppsala, Sweden.
authors: Olle
authors: O
authors: Korsgren
authors: Cytonome/ST, LLC, Boston, Massachusetts, USA.
authors: John C
authors: JC
authors: Sharpe
authors: Department of Immunology, Genetics and Pathology, Section of Transplantation Surgery, Uppsala University, Uppsala, Sweden; Department of Surgical Sciences, Section of Transplantation Surgery, Uppsala University, Uppsala, Sweden. Electronic address: david.berglund@igp.uu.se.
authors: David
authors: D
authors: Berglund
mesh_terms: D018791:CD4 Lymphocyte Count; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002470:Cell Survival; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D016219:Immunotherapy, Adoptive; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism; D053699:Interleukin-7 Receptor alpha Subunit / Q000378:metabolism; D007963:Leukocytes, Mononuclear; D044085:Microfluidics / Q000379:methods*; D050378:T-Lymphocytes, Regulatory / Q000166:cytology* / Q000378:metabolism; D013997:Time Factors
chemical_list: D053645:Interleukin-2 Receptor alpha Subunit; D053699:Interleukin-7 Receptor alpha Subunit
keywords: cell purification; cell separation; immunotherapy; regulatory T cells