改良血小板添加液低温液态保存血小板的形态及功能研究

Morphology and Function of Platelets Stored in Modified Platelet Additive Solution at Low Temperature

本研究探讨使用改良的血小板添加液替代70%自体血浆在低温(10℃)液态条件下对血小板的保存效果。采集6名供者单采血小板,每一名供者单采血小板平均分为3组,A组(常规保存组)加入100%血浆、22℃保存;B组(添加液10℃保存组)加入70%PAS-ⅢM/30%血浆、10℃保存;C组(冰冻保存组)加入100%血浆和海藻糖、-85℃保存;另设D组(血浆4℃保存组)加入100%血浆、4℃保存,作为扫描电子显微镜对照组。A、B组在第1、5、7、9天取样,C组在20天后复苏取样,分别检测CD62p、HSR、PAgT、LDH的变化。血浆常温组、添加液低温组于保存后第3、9天取样扫描电镜观察,血浆低温组于保存后第3天取样观察,冰冻组在20天后复苏取样观察,同时使用新鲜血小板作为对照。结果表明,保存期内随保存时间延长,A组和B组CD62p表达增加,HSR和最大聚集率降低;A组的LDH释放、CD62p表达、HSR、血小板最大聚集率与B组比较有显著统计学差异(p<0.05)。C组LDH释放明显较其他两组增多,但CD62p表达较少(p<0.05)。A、B组血小板形态保持较好,C组血小板形态保持差。结论:改良的PAS-ⅢM保存液能够替代血浆用于血小板的保存,在低温条件下能够很好地保护血小板的功能,保存效果优于血浆常温保存。

The aim of study was to evaluate the function of modified platelet additive solution （PAS-Ⅲ M） with trehalose as a substitute of plasma for the storage of platelet concentrates at low temperature （ 10℃ ）. Apheresis platelets from 6 donors were divided and added with different media （group A： 100% plasma; group B： 70% PAS-Ⅲ M/30% plasma; group C：100% plasma/trehalose）. Groups A, B,C were stored at 10℃, 22℃ and -85℃ separeately. In addition, group D （platelet concentrates stored with 100% plasma at 4℃ ） was set up as control group for scan electrornicroscopy. The samples of each platelets were collected on day 0, 1, 5, 7 and 9 after storage respectively, while samples of platelets stored at -85℃ （group C） were collected on day 20 after storage. CD62p, hypotonic shock response （ HSR）, platelet aggregation, lactic dehydrogenase （ LDH ） and morphology of platelets were evaluated. The results showed that the expressions of CD62p in groups A and B increased in a time-dependent manner, but HSR and platelet aggregations decreased. The expression of CD62p, LDH release, and platelet aggregation in group A were significant higher than that in group B （p 〈 0.05 ）. HSR in group A was significant lower than that in group B （p 〈 0. 05 ）. LDH release was significant high in samples of group C and the expression of CD62p was lower than that in other two groups （p 〈 0.05 ）. It is concluded that the protective effects of 70% PAS- Ⅲ M/30% plasma （ 10℃） and plasma platelets （22℃） on morphology of platelets are similar, but better than those of plasma platelets （4℃） and plasma/ trehalose （ -85℃ ）. In short, PAS-Ⅲ M serves as a good substitute of plasma for platelet storage, and protects the chilled platelets.