Research Article Platelet-Rich Plasma Obtained with Different Anticoagulants and Their Effect on Platelet Numbers and Mesenchymal Stromal Cells Behavior In Vitro Ronaldo José Farias Corrêa do Amaral, 1,2 Nemias Pereira da Silva, 1 Natália Ferreira Haddad, 1 Luana Siqueira Lopes, 1 Fábio Dias Ferreira, 1 Ricardo Bastos Filho, 3 Paola Alejandra Cappelletti, 1 Wallace de Mello, 1,4 Eric Cordeiro-Spinetti, 2 and Alex Balduino 1,2 1 Excellion Servic ¸os Biom´ edicos, Amil/UnitedHealth Group, 25651-000 Petr´ opolis, RJ, Brazil 2 Laborat´ orio de Biologia e Tecnologia Celular, Universidade Veiga de Almeida, 20270-150 Rio de Janeiro, RJ, Brazil 3 Universidade Federal Fluminense, 24033-900 Niter´ oi, RJ, Brazil 4 Centro Universit´ ario Celso Lisboa, 20950-091 Rio de Janeiro, RJ, Brazil Correspondence should be addressed to Ronaldo Jos´ e Farias Corrˆ ea do Amaral; ronaldojfca@gmail.com Received 8 January 2016; Revised 9 April 2016; Accepted 27 April 2016 Academic Editor: Jiabing Fan Copyright © 2016 Ronaldo Jos´ e Farias Corrˆ ea do Amaral et al. his is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. here are promising results in the use of platelet-rich plasma (PRP) for musculoskeletal tissue repair. However, the variability in the methodology for its obtaining may cause diferent and opposing indings in the literature. Particularly, the choice of the anticoag- ulant is the irst deinition to be made. In this work, blood was collected with sodium citrate (SC), ethylenediaminetetraacetic acid (EDTA), or anticoagulant citrate dextrose (ACD) solution A, as anticoagulants, prior to PRP obtaining. Hematological analysis and growth factors release quantiication were performed, and the efects on mesenchymal stromal cell (MSC) culture, such as cytotoxicity and cell proliferation (evaluated by MTT method) and gene expression, were evaluated. he use of EDTA resulted in higher platelet yield in whole blood; however, it induced an increase in the mean platelet volume (MPV) following the blood centrifugation steps for PRP obtaining. he use of SC and ACD resulted in higher induction of MSC proliferation. On the other hand, PRP obtained in SC presented the higher platelet recovery ater the blood irst centrifugation step and a minimal change in MSC gene expression. herefore, we suggest the use of SC as the anticoagulant for PRP obtaining. 1. Introduction Platelet-rich plasma (PRP) is a blood-derived product in which platelets are concentrated at least ive times in plasma above the baseline of that in the whole blood [1]. PRP is being investigated as an autologous product to improve tissue repair in diferent conditions and lesions, especially for muscu- loskeletal tissues, such as chondral lesions [2–4], tendinopa- thies [5–7], muscle strains [8, 9], and bone repair [10, 11]. Besides its clinical application, PRP may be an eicient sub- stitute to fetal bovine serum in cell culture [12–15]. Its thera- peutic potential is based mainly on the growth factors present in platelet’s alpha granules [16], such as transforming growth factor beta (TGF-) [17], vascular endothelial growth factor (VEGF) [18], and platelet-derived growth factor (PDGF) [19], which have already been demonstrated to play important roles in tissue repair. When platelets are concentrated and activated, it is expected that the concentration of the factors released reaches three to ive times of that found in the plasma [16]. he general methodology to obtain PRP involves the col- lection of whole blood with anticoagulants, followed by one or two centrifugation steps. Ater a irst low-speed centrifuga- tion, erythrocyte-free platelet concentrated plasma is recov- ered and submitted to high-speed centrifugation. Platelet- poor plasma is then discarded and the remaining platelet Hindawi Publishing Corporation Stem Cells International Volume 2016, Article ID 7414036, 11 pages http://dx.doi.org/10.1155/2016/7414036