A humanized system to expand in vitro amniotic fluid-derived stem cells intended for clinical application

Daniela Martinelli, Rui Cruz Pereira, Massimo Mogni, Roberto Benelli, Maddalena Mastrogiacomo, Domenico Coviello, Ranieri Cancedda, Chiara Gentili

Research output: Contribution to journalArticlepeer-review


Background aims 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.

Original languageEnglish
Pages (from-to)438-451
Number of pages14
Issue number3
Publication statusPublished - Mar 1 2016


  • Amniotic Fluid Stem Cells (AFSC)
  • fetal calf serum
  • platelet lysate
  • stem cells

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy
  • Oncology
  • Genetics(clinical)
  • Transplantation
  • Cancer Research
  • Cell Biology


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