Shin Hyeok-Soo, Woo Heung-Myong, Kang Byung-Jae
College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chuncheon, 24341, Republic of Korea.
BMC Vet Res. 2017 Jun 26;13(1):198. doi: 10.1186/s12917-017-1123-3.
Platelet-rich plasma (PRP) has been expected for regenerative medicine because of its growth factors. However, there is considerable variability in the recovery and yield of platelets and the concentration of growth factors in PRP preparations. The aim of this study was to identify optimal relative centrifugal force and spin time for the preparation of PRP from canine blood using a double-centrifugation tube method.
Whole blood samples were collected in citrate blood collection tubes from 12 healthy beagles. For the first centrifugation step, 10 different run conditions were compared to determine which condition produced optimal recovery of platelets. Once the optimal condition was identified, platelet-containing plasma prepared using that condition was subjected to a second centrifugation to pellet platelets. For the second centrifugation, 12 different run conditions were compared to identify the centrifugal force and spin time to produce maximal pellet recovery and concentration increase. Growth factor levels were estimated by using ELISA to measure platelet-derived growth factor-BB (PDGF-BB) concentrations in optimised CaCl-activated platelet fractions.
The highest platelet recovery rate and yield were obtained by first centrifuging whole blood at 1000 g for 5 min and then centrifuging the recovered platelet-enriched plasma at 1500 g for 15 min. This protocol recovered 80% of platelets from whole blood and increased platelet concentration six-fold and produced the highest concentration of PDGF-BB in activated fractions.
We have described an optimised double-centrifugation tube method for the preparation of PRP from canine blood. This optimised method does not require particularly expensive equipment or high technical ability and can readily be carried out in a veterinary clinical setting.
富含血小板血浆（PRP）因其生长因子而被寄予再生医学的厚望。然而，PRP制剂中血小板的回收率和产量以及生长因子的浓度存在很大差异。本研究的目的是使用双离心管法确定从犬类血液中制备PRP的最佳相对离心力和离心时间。
从12只健康的比格犬采集柠檬酸盐采血管中的全血样本。在第一次离心步骤中，比较了10种不同的运行条件，以确定哪种条件能实现血小板的最佳回收。一旦确定了最佳条件，使用该条件制备的含血小板血浆将进行第二次离心以沉淀血小板。在第二次离心中，比较了12种不同的运行条件，以确定产生最大沉淀回收率和浓度增加的离心力和离心时间。通过酶联免疫吸附测定法（ELISA）测量优化的氯化钙激活血小板组分中血小板衍生生长因子-BB（PDGF-BB）的浓度来估计生长因子水平。
通过先将全血在1000g下离心5分钟，然后将回收的富含血小板血浆在1500g下离心15分钟，可获得最高的血小板回收率和产量。该方案从全血中回收了80%的血小板，使血小板浓度增加了六倍，并在激活组分中产生了最高浓度的PDGF-BB。
我们描述了一种从犬类血液中制备PRP的优化双离心管法。这种优化方法不需要特别昂贵的设备或高技术能力，并且可以很容易地在兽医临床环境中进行。
