title: A humanized system to expand in vitro amniotic fluid-derived stem cells intended for clinical application.
abstract: OBJECTIVE
  The amniotic fluid is a new source of multipotent stem cells with therapeutic potential for human diseases. In agreement with the regulatory requirement to reduce and possibly to avoid animal-derived reagents in the culture of cells intended for cell therapy, bovine serum, the most common supplement in the culture medium, was replaced by human platelet-derived growth factors.
  
  
  METHODS
  We tested a new culture medium to expand monolayers of human amniotic fluid stem cells (hAFSC) for clinical use. The AFSC were isolated by c-Kit selection and expanded in media supplemented with either bovine serum or a human platelet lysate (Lyset).
  
  
  RESULTS
  We compared proliferation kinetics, colony-forming unit percentage, multilineage differentiation, immunophenotypic characterization and inhibition of peripheral blood mononuclear cell proliferation of the two AFSC cell cultures and we found no significant differences. Moreover, the karyotype analysis of the cells expanded in the presence of the platelet lysate did not present cytogenetic abnormalities and in vitro and in vivo studies revealed no cell tumorigenicity.
  
  
  CONCLUSIONS
  Platelet derivatives represent a rich source of growth factors that can play a safety role in the homeostasis, proliferation and remodeling of tissue healing. We propose human platelet extracts as a preferential alternative to animal serum for the expansion of stem cells for clinical applications.
authors: Laboratory of Regenerative Medicine, DIMES, IRCCS AOU San Martino-IST Largo Rosanna Benzi 10, University of Genova, Genova, Italy.
authors: Daniela
authors: D
authors: Martinelli
authors: Laboratory of Regenerative Medicine, DIMES, IRCCS AOU San Martino-IST Largo Rosanna Benzi 10, University of Genova, Genova, Italy.
authors: Rui Cruz
authors: RC
authors: Pereira
authors: Laboratory of Human Genetics, E.O. Ospedali Galliera, Genova, Italy.
authors: Massimo
authors: M
authors: Mogni
authors: Laboratory of Immunology, IRCCS AOU San Martino-IST Largo Rosanna Benzi 10, Genoa, Italy.
authors: Roberto
authors: R
authors: Benelli
authors: Laboratory of Regenerative Medicine, DIMES, IRCCS AOU San Martino-IST Largo Rosanna Benzi 10, University of Genova, Genova, Italy.
authors: Maddalena
authors: M
authors: Mastrogiacomo
authors: Laboratory of Human Genetics, E.O. Ospedali Galliera, Genova, Italy.
authors: Domenico
authors: D
authors: Coviello
authors: Laboratory of Regenerative Medicine, DIMES, IRCCS AOU San Martino-IST Largo Rosanna Benzi 10, University of Genova, Genova, Italy.
authors: Ranieri
authors: R
authors: Cancedda
authors: Laboratory of Regenerative Medicine, DIMES, IRCCS AOU San Martino-IST Largo Rosanna Benzi 10, University of Genova, Genova, Italy. Electronic address: chiara.gentili@unige.it.
authors: Chiara
authors: C
authors: Gentili
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D003470:Culture Media / Q000378:metabolism / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000502:physiology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000502:physiology; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D039902:Multipotent Stem Cells / Q000166:cytology / Q000502:physiology; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects
chemical_list: D003470:Culture Media
keywords: Amniotic Fluid Stem Cells (AFSC); fetal calf serum; platelet lysate; stem cells