title: Does Freeze-Thawing Influence the Effects of Platelet Concentrates? An In Vitro Study on Human Adipose-Derived Stem Cells.
abstract: Human adipose-derived stem cells (hASCs) have been proposed as a possible therapy for tissue regeneration in aesthetic, plastic, and reconstructive surgery. Today, platelet concentrates are used in a wide range of disciplines, but their storage has become a controversial aspect. The purpose of this in vitro study was to evaluate the effect of plasma rich in growth factors (PRGF), after a freeze-thawing cycle, on the proliferation and biological activity of progenitor cells involved in soft tissue healing. Different formulations of activated PRGF were added to hASCs cultured in serum-free medium. Cell proliferation was assessed by MTT test and cell count up to 7 and 12-day incubation. Osteo-differentiation ability of hASCs was also tested after 7 and 14-day incubation by alkaline phosphatase assay. The effects of 4 PRGF preparations (fresh/frozen and with/without platelets) were compared with corresponding formulations of plasma poor in growth factors and with standard medium. hASCs cultured in the presence of platelet concentrates increased proliferation rate with respect to cells grown in standard medium without significant differences among all the tested plasma formulations on cell viability up to 12 days of culture. PRGF activity is preserved after cryopreservation and platelet-rich preparations promoted osteo-differentiation of hASCs at day 7. In conclusion, PRGF supports the proliferation and the differentiation of progenitor cells in vitro also when applied after cryopreservation. Platelet concentrates, either alone or in combination with mesenchymal stem cells, might be a valuable tool in the field of tissue regeneration.
authors.affiliation: *Department of Biomedical Surgical and Dental Sciences, Università degli Studi di Milano, Milan, Italy †IRCCS Galeazzi Orthopaedic Institute, Milan, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Ceci
authors.forename: Stefania
authors.initials: S
authors.lastname: Niada
authors.forename: Massimo
authors.initials: M
authors.lastname: Del Fabbro
authors.forename: Alessandra
authors.initials: A
authors.lastname: Lolato
authors.forename: Silvio
authors.initials: S
authors.lastname: Taschieri
authors.forename: Chiara
authors.initials: C
authors.lastname: Giannasi
authors.forename: Anna Teresa
authors.initials: AT
authors.lastname: Brini
mesh_terms: D017667:Adipocytes / Q000166:cytology*; D000328:Adult; D001792:Blood Platelets / Q000166:cytology*; D002452:Cell Count; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D005615:Freezing; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*; D055815:Young Adult