Autologous Non-Activated Platelet Rich Plasma (PRP) a Proliferation Promoting Substitute for Xenogenic Culture Media

Adipose derived Mesenchymal stem cells (AT-MSC) are promising cells for cell therapy and tissue engineering. However, their expansion needs in vitro culture. Actually non-autologous culture media (e.g fetal bovine serum (FBS), plasma from blood bank) that brings potential immunological reactions and are used. In this study we investigate the effects of non-activated autologous platelet-rich plasma (nPRP) and autologous thrombin activated platelet rich plasma (tPRP) on AT-MSCs proliferation, as a FBS substitute.

Methods.

AT-MSCs of 10 donors were isolated and cultured for 10 days under different conditions: (a) 10% FBS as control, (b) 1%, (c) 5%, (d) 10%, (e) 20%, (f) 40% and (g) 60% of tPRP or nPRP. Platelet viability was assessed by Calcein. The proliferation capacity of nPRP and FBS cultured cells was quantified by EDU assay. MSCs phenotype markers were analyzed by FACS calibur. Differentiation was proved by corresponding staining.

Results.

57% of platelets were viable until 10 days of culture. PRP increased AT-MSC proliferation dose dependently: 20% nPRP provided a higher cell proliferation rate than tPRP and FBS while MSCs phenotype and differentiation potency were not modified.

Conclusion.

 20% nPRP could serve as a potent substitute for non-autologous media in human cell therapies and tissue engineering researches based on GMP protocols.