血液实验教学中血浆制备的条件优化及储存时间对实验效果的影响

摘要

目的：对生理学血液实验教学中富血小板血浆（ platelet-rich plasma，PRP）及少血小板血浆（platelet-poor plasma，PPP）制备条件进行优化，并明确血浆储存时间对实验效果的影响。方法：比较家兔麻醉状态下不同采血方法、家兔有无麻醉、采血后血液置于不同材质的试管以及不同离心参数等因素对 PRP 及 PPP 凝血时间以及得率的影响；并且测定储存时间对制备的血浆凝固时间的影响。结果：（1）家兔麻醉状态下，三种采血法（头皮针颈动脉采血、颈动脉放血、心脏采血）无明显差异；（2）采用心脏采血方法，在家兔清醒状态下采血所得 PRP 凝固时间比麻醉状态下的时间更短（P ﹤0.001）；（3）玻璃离心管作为容器所得 PRP 和 PPP 的凝固时间分别较塑料离心管的短（P ﹤0.001），且塑料离心管所得 PPP 得率高于玻璃采血管（P ﹤0.01）；（4）三种离心参数制备 PRP，以Aghaloo 法 PRP 的凝固时间最长（P ﹤0.001），本科室传统离心法与 Landesberg 法的凝固时间无统计学差异；Land-esberg 法和 Aghaloo 法的 PRP 得率无差异，但较本科室传统离心法高（P ﹤0.001）；（5）PRP 和 PPP 于-20℃储存1~360 d 后，对血浆凝固时间无明显影响。结论：PRP 的制备以家兔在清醒状态下、心脏采血、采用玻璃离心管、以 Landesberg 法离心，血浆得率高、凝固时间短；PPP 的制备采用塑料离心管，以1500× g 离心30 min，血浆得率高、凝固时间长。将所得血浆分装，-20℃保存，血小板活性可保持一年。%Objective:To study the optimization of the preparation of PRP and PPP and the effect of plasma′s storage time on clotting time in physiology blood experimental teaching. Methods:The clotting time and the yield of PRP and PPP were determined by comparing blood samples,which were sampled from anaesthetized rabbits in different ways and unan-aesthetized rabbits and were stored in different kind of tube material and different parameters of centrifugal. At the same time,the effect of plasma′s storage time on clotting time was measured. Results:(1)In anaesthetized rabbits,there was no significant difference of clotting time in the three different blood sampling methods,which collected blood from carotid artery by scalp acupuncture,from carotid artery by intubation,and from heart by punctures.(2)By taking the method of heart punctures to collect blood,the clotting time of PRP from the conscious rabbit was shorter than from the anaesthe-tized rabbit(P ﹤ 0. 001).(3)The clotting time of PRP and PPP prepared in the glass tubes were shorter than in the plastic tubes(P ﹤ 0. 001),and the yield of PPP prepared in the plastic tubes were more than in the glass tubes(P ﹤0. 01).(4)The clotting time of PRP prepared by Aghaloo method was the longest(P ﹤ 0. 001),while there was no sta-tistical difference between the traditional centrifugal method and Landesberg method. But the yield of PRP prepared by Landesberg method and Aghaloo method was undifferentiated,which was higher than the traditional centrifugal method(P﹤ 0. 001).(5)After being stored at - 20 ℃ for 1 ~ 360 d,the clotting time of PRP and PPP was not affected. Conclu-sion:It is better to prepare PRP in conscious rabbit,to collect blood by heart punctures,to use glass tube and be pre-pared by Landesberg method,whereas the preparation of PPP is better by using plastic tube,and being centrifuged in 1 500 × g 30 mintues . Even more,the platelet activity can be kept for one year if the plasma is divided and stored at- 20 ℃ .