title: Inactivated human platelet lysate with psoralen: a new perspective for mesenchymal stromal cell production in Good Manufacturing Practice conditions.
abstract: OBJECTIVE
  Mesenchymal stromal cells (MSC) are ideal candidates for regenerative and immunomodulatory therapies. The use of xenogeneic protein-free Good Manufacturing Practice-compliant growth media is a prerequisite for clinical MSC isolation and expansion. Human platelet lysate (HPL) has been efficiently implemented into MSC clinical manufacturing as a substitute for fetal bovine serum (FBS). Because the use of human-derived blood materials alleviates immunologic risks but not the transmission of blood-borne viruses, the aim of our study was to test an even safer alternative than HPL to FBS: HPL subjected to pathogen inactivation by psoralen (iHPL).
  
  
  METHODS
  Bone marrow samples were plated and expanded in α-minimum essential medium with 10% of three culture supplements: HPL, iHPL and FBS, at the same time. MSC morphology, growth and immunophenotype were analyzed at each passage. Karyotype, tumorigenicity and sterility were analyzed at the third passage. Statistical analyses were performed.
  
  
  RESULTS
  The MSCs cultivated in the three different culture conditions showed no significant differences in terms of fibroblast colony-forming unit number, immunophenotype or in their multipotent capacity. Conversely, the HPL/iHPL-MSCs were smaller, more numerous, had a higher proliferative potential and showed a higher Oct-3/4 and NANOG protein expression than did FBS-MSCs. Although HPL/iHPL-MSCs exhibit characteristics that may be attributable to a higher primitive stemness than FBS-MSCs, no tumorigenic mutations or karyotype modifications were observed.
  
  
  CONCLUSIONS
  We demonstrated that iHPL is safer than HPL and represents a good, Good Manufacturing Practice-compliant alternative to FBS for MSC clinical production that is even more advantageous in terms of cellular growth and stemness.
authors: Pediatric Onco-Hematology, Stem Cell Transplantation and Cellular Therapy Division, City of Science and Health of Turin, Regina Margherita Children's Hospital, Turin, Italy.
authors: Sara
authors: S
authors: Castiglia
authors: Pediatric Onco-Hematology, Stem Cell Transplantation and Cellular Therapy Division, City of Science and Health of Turin, Regina Margherita Children's Hospital, Turin, Italy; Department of Public Health and Pediatrics, University of Turin, Turin, Italy. Electronic address: katia.mareschi@unito.it.
authors: Katia
authors: K
authors: Mareschi
authors: Blood Component Production and Validation Center, City of Science and Health of Turin, S. Anna Hospital, Turin, Italy.
authors: Luciana
authors: L
authors: Labanca
authors: Blood Component Production and Validation Center, City of Science and Health of Turin, S. Anna Hospital, Turin, Italy.
authors: Graziella
authors: G
authors: Lucania
authors: Pediatric Onco-Hematology, Stem Cell Transplantation and Cellular Therapy Division, City of Science and Health of Turin, Regina Margherita Children's Hospital, Turin, Italy.
authors: Marco
authors: M
authors: Leone
authors: Pediatric Onco-Hematology, Stem Cell Transplantation and Cellular Therapy Division, City of Science and Health of Turin, Regina Margherita Children's Hospital, Turin, Italy.
authors: Fiorella
authors: F
authors: Sanavio
authors: Pediatric Onco-Hematology, Stem Cell Transplantation and Cellular Therapy Division, City of Science and Health of Turin, Regina Margherita Children's Hospital, Turin, Italy.
authors: Laura
authors: L
authors: Castello
authors: Pediatric Onco-Hematology, Stem Cell Transplantation and Cellular Therapy Division, City of Science and Health of Turin, Regina Margherita Children's Hospital, Turin, Italy.
authors: Deborah
authors: D
authors: Rustichelli
authors: Pediatric Onco-Hematology, Stem Cell Transplantation and Cellular Therapy Division, City of Science and Health of Turin, Regina Margherita Children's Hospital, Turin, Italy.
authors: Elena
authors: E
authors: Signorino
authors: Pediatric Onco-Hematology, Stem Cell Transplantation and Cellular Therapy Division, City of Science and Health of Turin, Regina Margherita Children's Hospital, Turin, Italy.
authors: Monica
authors: M
authors: Gunetti
authors: Department of Public Health and Pediatrics, University of Turin, Turin, Italy.
authors: Massimiliano
authors: M
authors: Bergallo
authors: Blood Component Production and Validation Center, City of Science and Health of Turin, S. Anna Hospital, Turin, Italy.
authors: Anna Maria
authors: AM
authors: Bordiga
authors: Pediatric Onco-Hematology, Stem Cell Transplantation and Cellular Therapy Division, City of Science and Health of Turin, Regina Margherita Children's Hospital, Turin, Italy; Department of Public Health and Pediatrics, University of Turin, Turin, Italy.
authors: Ivana
authors: I
authors: Ferrero
authors: Pediatric Onco-Hematology, Stem Cell Transplantation and Cellular Therapy Division, City of Science and Health of Turin, Regina Margherita Children's Hospital, Turin, Italy.
authors: Franca
authors: F
authors: Fagioli
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000166:cytology*; D002417:Cattle; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics; D002457:Cell Extracts; D049109:Cell Proliferation / Q000235:genetics; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology*
chemical_list: D002457:Cell Extracts
keywords: Good Manufacturing Practice; human platelet lysate; inactivation; mesenchymal stromal cells; psoralen