OBJECTIVE: 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 (PCs) are used in a wide range of disciplines, but their storage has become a controversial aspect. The aim of this in vitro study was to assess the effect of the platelet concentrate Plasma Rich in Growth Factors (PRGF), after a freeze-thawing cycle, on the proliferation and biological activity of progenitor cells involved in soft tissue regeneration, using PRGF as substitute for fetal bovine serum (FBS). MATERIALS AND METHODS: Four different formulations of activated PRGF (fresh PRGF with platelets; frozen PRGF with platelets; fresh PRGF without platelets and frozen PRGF without platelets), obtained from human donors, were added to human adipose derived stem cells (hASCs) cultured in serum-free medium. Other four formulations were prepared in which Plasma Poor in Growth Factors (PPGF) was used in place of PRGF to evaluate the relative contribution of plasmatic factors and factors released by platelets on the observed effects. The effects of the eight above preparations were compared with standard medium. Cell proliferation was assessed by MTT test and cell count up to 7 and 12-days incubation. Osteo-differentiation ability of hASCs was also determined after 7 and 14 days-incubation by Alkaline Phosphatase assay (ALP). Statistical analysis was performed using one-way analysis of variance and differences were considered significant at p<0.05. RESULTS: After a short adaptation period, hASCs cultured in the presence of PRGF induces an increased proliferation rate respect to cells grown in standard medium. No significant difference in the observed effect on cell viability was found among all the tested plasma formulations up to 12 days of culture. Regarding cell count in all the tested groups, the number of hASCs collected after 12 days was about 10 times higher than the control. No donor effect was observed. Plasma preparations promoted osteo-differentiation of hASCs, with a peak of activity at day 7. CONCLUSION: PRGF preparations exert a positive effect on the proliferation and the differentiation of progenitor cells in vitro. PRGF activity is preserved after cryopreservation. Plasma Poor in Growth Factors appears to have the ability to promote wound healing as well. Platelet concentrates in combination with human adipose-derived stem cells might be a valuable tool in the field of tissue regeneration.
|Titolo:||Does freeze-thawing influence the effects of platelet concentrates? : an in vitro study on human adipose-derived stem cells|
CECI, CATERINA (Primo)
NIADA, STEFANIA (Secondo)
|Data di pubblicazione:||9-ott-2015|
|Settore Scientifico Disciplinare:||Settore BIO/13 - Biologia Applicata|
|Citazione:||Does freeze-thawing influence the effects of platelet concentrates? : an in vitro study on human adipose-derived stem cells / C. Ceci, S. Niada, M. Del Fabbro, A. Lolato, S. Taschieri, C. Giannasi, A.T. Brini. ((Intervento presentato al convegno GISM Annual Meeting tenutosi a Brescia nel 2015.|
|Appare nelle tipologie:||01 - Articolo su periodico|