ABSTRACT
BACKGROUND: Adipose-derived stem cells (ASCs) are a powerful tool for cosmetic surgery and regenerative medicine. The use of autologous platelet rich plasma (PRP), particularly in combination with ASC-based therapy, has significantly expanded in recent years. Unfortunately, the mechanisms and optimal dosing responsible for the beneficial effects of PRP remain poorly understood. Here we investigate the effect of PRP on ASC growth and differentiation. OBJECTIVES: To assess the impact of different PRP feeding and cryopreservation protocols on ASC isolation, expansion, and differentiation. METHODS: Human PRP was isolated using the Magellan System (Arteriocyte). Fresh PRP (fPRP), flash frozen PRP (ffPRP), and cryopreserved PRP (cPRP) were added to human ASCs isolated from healthy patients. A panel of PRP supplementation protocols was analyzed for ASC adherence, proliferation, and osteogenesis. RESULTS: The fresh and cryopreserved PRP groups demonstrated reduced cell adherence compared to control (non-PRP) groups (P < 0.001), while the flash frozen PRP groups showed cell adherence equivalent to or better than controls. After 7 days of growth, ASC populations for fPRP and ffPRP Single Administration protocols were significantly higher than other feeding protocols and controls. This benefit was lost in cPRP groups. Optimized ffPRP protocols showed potential for spontaneous osteogenesis. CONCLUSIONS: Addition of ffPRP improves initial ASC adherence while a single administration of either fresh or flash frozen PRP without additional cell manipulation significantly augments subsequent ASC proliferation. The potential for spontaneous osteogenic differentiation upon PRP exposure invokes the need for additional molecular studies of PRP activity prior to further expansion to clinical applications.