【摘要】 目的研究人血小板冻干保存前负载海藻糖至细胞内过程中DMSO的作用,优化血小板负载缓冲液配方。方法实验设对照组(负载缓冲液中未添加DMSO)和实验组(负载缓冲液中添加DMSO),使用流式细胞仪检测血小板负载海藻糖4h后膜表面糖蛋白分子CD62p、PAC-1的表达,通过激光共聚焦荧光显微镜观察血小板负载荧光物质LYCH4h后其胞内LYCH的分布,同时检测实验组和对照组负载海藻糖4h后血小板平均体积(MPV)。结果两组结果比较,实验组CD62p、PAC-1的表达显著低于对照组(P<0.05),实验组LYCH均匀分布在血小板胞质中,而对照组LYCH大部分分布在几个有限细胞器内,两组MPV未有显著性差别(P>0.05)。结论DMSO在血小板负载海藻糖过程中可有效抑制血小板体外激活,并使胞质内海藻糖均匀分布,更好发挥海藻糖对细胞的保护作用。更多还原

【Abstract】 Objective To study the effect of DMSO on platelets protection during the period of loading trehalose before lyophilization, and to optimize the method of loading buffer.Methods The samples were divided into two groups: the control group (without adding DMSO to the loading buffer), and the test group (adding DMSO to the loading buffer). Flow cytometry was used to assay the expressions of glycoprotein CD62p and PAC-1, and the fluorescense microscope was used to observe the uptake of LYCH and its intracellular distribution 4 hours after incubation. The values of MPV between the two trehalose loaded groups were assayed by cytometer. Results Compared to the control group, the expressions of CD62p and PAC-1 were significantly lower (P<0.05). LYCH of test group was distributed homogeneously in the cytoplasm of platelets; however, in the control platelets, LYCH was distributed mostly in a few organelles. Conclusion DMSO can effectively inhibit the platelets activation 4 hours after incubation, contribute to more homogeneous distribution of trehalose in the cytoplasm, and enhance trehalose protection of platelets.更多还原