title: Overcoming the bottleneck of platelet lysate supply in large-scale clinical expansion of adipose-derived stem cells: A comparison of fresh versus three types of platelet lysates from outdated buffy coat-derived platelet concentrates.
abstract: Platelet lysates (PL) represent a promising replacement for xenogenic growth supplement for adipose-derived stem cell (ASC) expansions. However, fresh platelets from human blood donors are not clinically feasible for large-scale cell expansion based on their limited supply. Therefore, we tested PLs prepared via three methods from outdated buffy coat-derived platelet concentrates (PCs) to establish an efficient and feasible expansion of ASCs for clinical use.
  
  
  
  PLs were prepared by the freeze-thaw method from freshly drawn platelets or from outdated buffy coat-derived PCs stored in the platelet additive solution, InterSol. Three types of PLs were prepared from outdated PCs with platelets suspended in either (1) InterSol (not manipulated), (2) InterSol + supplemented with plasma or (3) plasma alone (InterSol removed). Using these PLs, we compared ASC population doubling time, cell yield, differentiation potential and cell surface markers. Gene expression profiles were analyzed using microarray assays, and growth factor concentrations in the cell culture medium were measured using enzyme-linked immunosorbent assay (ELISA).
  
  
  
  Of the three PL compositions produced from outdated PCs, removal of Intersol and resuspension in plasma prior to the first freezing process was overall the best. This specific outdated PL induced ASC growth kinetics, surface markers, plastic adherence and differentiation potentials comparable with PL from fresh platelets. ASCs expanded in PL from fresh versus outdated PCs exhibited different expressions of 17 overlapping genes, of which 10 were involved in cellular proliferation, although not significantly reflected by cell growth. Only minor differences in growth factor turnover were observed.
  
  
  
  PLs from outdated platelets may be an efficient and reliable source of human growth supplement allowing for large-scale ASC expansion for clinical use.
authors.affiliation: Department of Plastic Surgery, Breast Surgery & Burns, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark. Electronic address: peter.viktor.glovinski.01@regionh.dk.
authors.forename: Peter V
authors.initials: PV
authors.lastname: Glovinski
authors.affiliation: Department of Plastic Surgery, Breast Surgery & Burns, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark.
authors.forename: Mikkel
authors.initials: M
authors.lastname: Herly
authors.affiliation: Cardiology Stem Cell Laboratory, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark.
authors.forename: Anders B
authors.initials: AB
authors.lastname: Mathiasen
authors.affiliation: Cell Therapy Facility, The Blood Bank, Department of Clinical Immunology, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark.
authors.forename: Jesper D
authors.initials: JD
authors.lastname: Svalgaard
authors.affiliation: Center for Genomic Medicine, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark.
authors.forename: Rehannah
authors.initials: R
authors.lastname: Borup
authors.affiliation: Department of Pathology, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark.
authors.forename: Maj-Lis M
authors.initials: MM
authors.lastname: Talman
authors.affiliation: Department of Plastic Surgery, Breast Surgery & Burns, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark.
authors.forename: Jens J
authors.initials: JJ
authors.lastname: Elberg
authors.affiliation: Department of Plastic Surgery, Breast Surgery & Burns, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark.
authors.forename: Stig-Frederik T
authors.initials: ST
authors.lastname: Kølle
authors.affiliation: Department of Plastic Surgery, Breast Surgery & Burns, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark.
authors.forename: Krzysztof T
authors.initials: KT
authors.lastname: Drzewiecki
authors.affiliation: Cell Therapy Facility, The Blood Bank, Department of Clinical Immunology, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark.
authors.forename: Anne
authors.initials: A
authors.lastname: Fischer-Nielsen
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology* / Q000502:physiology; D058621:Blood Buffy Coat / Q000166:cytology* / Q000637:transplantation; D001792:Blood Platelets / Q000737:chemistry / Q000166:cytology*; D001793:Blood Preservation / Q000379:methods*; D001800:Blood Specimen Collection / Q000379:methods; D018929:Cell Culture Techniques / Q000379:methods*; D002457:Cell Extracts / Q000600:supply & distribution*; D049109:Cell Proliferation; D002469:Cell Separation; D003470:Culture Media / Q000378:metabolism; D005260:Female; D005615:Freezing; D006801:Humans; D010949:Plasma / Q000166:cytology; D017713:Platelet Transfusion / Q000379:methods; D053657:Platelet-Rich Plasma / Q000166:cytology; D012034:Refrigeration; D013997:Time Factors
chemical_list: D002457:Cell Extracts; D003470:Culture Media
keywords: adipose-derived stem cells; cell proliferation; cell therapy; gene expression; growth factors; platelet lysate