人血小板冻干前海藻糖负载技术的优化

本研究以建立血小板负载海藻糖基本技术为基础，进一步优化海藻糖负载技术，包括负载温度、负载时间、海藻糖负载浓度和负载溶液。通过比较缓冲液和血浆负载效果，比较生理温度（37℃）和相变温度（16℃）下血小板胞内海藻糖浓度与负载时间曲线、血小板平均体积（MPV）与负载时间、海藻糖负载浓度的曲线、计算海藻糖的负载率，优选负载率较高的负载溶液、负载温度、负载时间和海藻糖浓度等血小板海藻糖负载技术参数。结果表明：37℃血浆负载率高于缓冲液负载率，且血浆负载4小时，在37℃海藻糖负载率可高达19.51％，明显高于16℃的负载率6、16％；血小板MPV在16℃比在37℃明显增大43、2％，随海藻糖负载时间（1—4小时）延长和负载浓度（0—100mmol／L）增加未见明显改变；血小板MPV在37℃与海藻糖负载时间和负载浓度呈正相关性，且海藻糖负载时间与负载浓度存在协同效应，海藻糖负载浓度高于50mmol／L，MPV随浓度（0—100mmol／L）、负载时间（1—4小时）增加而增大。结论：在原血浆中负载海藻糖、温度37℃、时间4小时、浓度低于50mmot／L可以作为优化后血小板海藻糖负载技术基本参数，负载浓度需根据冻干保存需要进行调整。

Optimization on Trehalose Loading Technique as Protective Conditioning for Lyophilization of Human Platelets

This study was aimed to further optimize trehalose loading technique including loading temperature, loading time, loading solution and loading concentration of trehalose, based on the established parameters. Loading efficiency in plasma was compared with that in buffer at 37 degrees C; the curves of intracellular trehalose concentration versus loading time at 37 degrees C and 16 degrees C were measured; curves of mean platelet volume (MPV) versus loading time and loading concentration were investigated and compared. According to results obtained, the loaing time, loading temperature, loading solution and trehalose concentration were ascertained for high loading efficiency of trehalose into human platelet. The results showed that the loading efficiency in plasma was markedly higher than that in buffer at 37 degrees C, the loading efficiency in plasma at 37 degrees C was significantly higher than that at 16 degrees C and reached 19.51% after loading for 4 hours, but 6.16% at 16 degrees C. MPV at 16 degrees C was increased by 43.2% than that at 37 degrees C, but had no distinct changes with loading time and loading concentration. In loading at 37 degrees C, MPV increased with loading time and loading concentration positively. Loading time and loading concentration displayed synergetic effect on MPV. MPV increased with loading time and concentration while trehalose loading concentration was above 50 mmol/L. It is concluded that the optimization parameters of trehalose loading technique are 37 degrees C (temperature), 4 hours (leading time), plasma (loading solution), 50 mmol/L (feasible trehalose concentration). The trehalose concentration can be adjusted to meet the requirement of lyophilization.