Objective: To optimize the single centrifugation preparation protocol of rat platelet-rich plasma (PRP).
Methods: The arterial blood of rats obtained by carotid artery intubation was collected by heparin sodium anticoagulant tubes, and then the blood divided into sterile EP tubes, adjusting the red blood cell concentration with normal saline, while rat PRP was prepared by centrifugation under different conditions (the centrifugal force was 200×g-240×g, and the centrifugal time was 8-12 min). Subsequently, the blood cell count and quality evaluation of anticoagulat whole blood and PRP were performed by hematology analyzer and flow cytometry, respectively, and the differences between different groups were compared.
Results: The red blood cell concentration to (5.5-6.5)×1012/L after anticoagulation of rat whole blood was good for PRP extraction. When the blood samples was centrifuged at 220×g for 10 min, the platelet recovery rate was the highest［(53.52±0.63)%］. The level of apoptosis and activation of plateles in PRP were not significantly different compared to whole blood(P>0.05), and the release level of growth factor was significantly increased(P<0.05).
Conclusion: It is a key to improve the PRP extraction efficiency by reducing the amount of mixed red blood cell, and this study successfully modified the preparation method of rat PRP, with platelets high recovery rate and stable quality.
题目: 大鼠富血小板血浆制备方法的改良及质量检测.
目的: 优化一次离心法制备大鼠富血小板血浆（platelet-rich plasma, PRP）的实验方案.
方法: 用肝素钠抗凝管收集经颈动脉插管获取的大鼠动脉血后分装入无菌EP管中，使用生理盐水调整红细胞浓度后，于不同条件下（200、220、240×g 离心8、10、12 min等）制备大鼠PRP，随后利用血细胞分析仪及流式细胞仪分别对抗凝全血及PRP进行细胞计数和质量评价，比较不同组间的差异.
结果: 大鼠全血抗凝后调整红细胞浓度至（5.5-6.5）×1012/L时有利于PRP的提取。当以220×g离心10 min时，血小板回收率最高［（53.52±0.63）%］。且提取出的PRP凋亡情况和血小板活化水平与全血无明显差异（P>0.05）。提取的PRP中生长因子释放水平较全血显著升高（P<0.05）.
结论: 降低红细胞混入量是提高PRP提取效率的关键，本研究成功改良了大鼠PRP的制备方法，血小板的回收率高且质量稳定.
Keywords: centrifugal conditions; platelet-rich plasma; rat; the amount of red blood cell mixing.