摘要
目的探讨冷冻干燥法对血小板的细胞形态、超微结构和细胞凋亡的影响。方法以新鲜血小板为对照,以保存30d后再水化的冻干血小板为实验对象,分别在倒置相差显微镜和透射电镜下观察血小板的细胞形态和超微结构,用流式细胞仪(FCM)检测血小板凋亡率,并以1×103U/L凝血酶为诱导剂检测再水化血小板的聚集活性。结果与新鲜血小板相比,冻干再水化后的血小板在形态上为圆盘状,未见有明显的细胞聚集,仅细胞透光性相对较弱;超微结构上也无明显变化,其内部基本结构清晰可辨,细胞膜、细胞器及分泌颗粒的包膜保持完整。新鲜血小板的细胞早期凋亡率为(0.44±0.19)%,显著低于冻干血小板的细胞凋亡率(3.25±1.68)%(P<0.05)。冻干再水化血小板的最大聚集率为(93.28±2.3)%。结论冻干再水化的血小板形态结构完整,细胞内超微结构完好,为本实验室成功建立血小板冻干保存方法提供了依据。
Objective To evaluate the effect of lyophilization on the morphological feature, ultrastructure and apoptosis of platelets. Methods The lyophilized platelets were rehydrated after being preserved 30 days. The change of morphology and ultrastructure of platelets was observed under the light microscope and transmission electron microscope （TEM）. The survival rate and apoptosis rate of platelets were analyzed by flow cytometry （FCM）. The aggregation rates of rehydrated lyophilized platelets were measured by using 1 U/mL thrombin. Results The morphology of rehydrated lyophilized platelets keeps normal. There was no evident cell aggregation in rehydrated lyophilized platelets, but the refractive index was relative low as compared with fresh platelets. The result of ultrastructure observation showed that the intracellular elementary structures of rehydrated lyophilized platelets were clear and the cell membrane keeps intact. According to the result of FCM analysis, the apoptosis rate of the fresh platelets and lyophilized platelets was（0. 44 ± 0. 19）% and （3.25 ± 1.68）% respectively, and there was a significant difference between the two groups （ P 〈 0.05 ）. The maximum aggregation rates of rehydrated lyophilized platelets were （ 93.28 ± 2.3 ） %. Conclusion The rehydrated lyophilized platelets keepintact cellular structure. The result supports method building of lyophilized platelet in our lab.
出处
《基础医学与临床》
CSCD
北大核心
2008年第11期1169-1173,共5页
Basic and Clinical Medicine
基金
卫生部科学研究基金省部共建项目计划(WKJ2005-2-037)
浙江省卫生高层次创新人才培养工程项目
关键词
冻干血小板
再水化
超微结构
细胞凋亡
lyophilized platelet
rehydrated
ultrastructure
apoptosis
