ID: 2214

• Title:
  Selective Pulse electric fields for valorization of Platelets Releasates towards biomedical potential product
  
  Authors:
  

  Salvador, Daniela1.2, Almeida, Henrique 1.2, Rego, Duarte5, Mendonça, Pedro3, Sousa, Ana P. 3, Redondo, Luís 4, Serra, Margarida 1,2 
  1.iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.
  2.ITQB-NOVA, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal
  3.IPST, Instituto Português do Sangue e da Transplantação, Lisboa, Portugal
  4.ISEL, Instituto Superior de Engenharia de Lisboa, Portugal
  5.EPS, Energy Pulse Systems, P

  
  
  Abstract:
  Mesenchymal stem/stromal cells (MSC) have an important role during tissue regeneration, not only because of their ability to differentiate into mesodermal lineage cells but also due to their rich secretome. Tissue regeneration is a physiological process intrinsic to several organs, however some tissue have a very limited regenerative capacity, such as heart, liver and central nervous system. Clinical trials using MSC are exponential increasing and showing promising results in the regenerative field. Nevertheless, animal derived components are still being used in advanced MSC manufacturing, such as the use of Fetal Bovine Serum (FBS) as media supplement. Platelets contain a wide range of bioactive factors, which are released upon platelet stimulation. Studies have shown that human platelet derivatives can be used as media supplementation during MSC production. Thus, the aim of this study is to develop a platelet derived xeno-free media supplement, through the application of Pulse Electric Field (PEF) to platelets concentrates (PC) with no therapeutic value. Posteriorly, evaluate the potential of these Platelet Releaseates obtained after PEF application (PR PEF) to sustain Bone Marrow-MSC (BM-MSC) self-renew and differentiation.
  
  In the current study, we assessed the impact of key PEF parameters, namely pulse width, electric field and pulse number, on platelet activation and growth factor release, and compare it to platelet lysates (PL) obtained by freeze and thaw cycles. We observed that PEF application to expired PC induces platelet activation (CD62P positive platelets) and leads to a controlled release of growth factors from the platelets. PR PEF features were dependent on PEF parameters applied. PEF application induces the release of PDGF, FGF, IGF and TGF-β from platelets, with the last one attaining similar levels to PL. A variability on growth factor levels obtained after PEF application in comparison to PL, indicates that growth factor might be localized differently in the platelet and that PEF is able to differently affect the platelet granules.
  
  Moreover, our data showed that BM-MSC were expanded in presence of PR PEF, with highest levels of TGF-β1 and FGF, retain their MSC features, namely the cell-surface markers, the ability to proliferate and differentiate into osteogenic, adipogenic and chondrogenic lineages.
  
  In conclusion, the application of PEF parameters can regulate the levels of bioactive components released by platelets obtained from expired PC and tuned PR features. These PR PEF are a suitable alternative to FBS used in MSC production.
  
  Further studies are required to characterize the secretome of MSC expanded in media supplemented with PR obtained following specific PEF parameters application.
  
  
  
  Keywords:
  
  
  Refs:
  
  
  Topic 1:
  12. Biomedical applications
  
  Topic 2:
  1. Biological responses (molecular, subcellular, cellular and intercellular)