A Study on Trehalose Loading Technique of Human Platelets before Freezing-dried / 中国医师杂志

ABSTRACT

Objective To establish the technique of loading trehalose into human platelets including temperature, time and concentration of trehalose loading. Methods Intracellular trehalose concentration versus loading time curves at 37℃ and 16℃, mean plateletee volume(MPV) versus loading time curves, and loading concentration curves were investigated and compared. Then loading time and temperature for high loading efficiency of trehalose into human platelets were ascertained. Intracellular trehalose versus trehalose loading concentration curves were made, and loading efficiency was caculated. Results The trehalose loading efficiency of 4 hours at 37℃ was significantly higher than that at 16℃ (11.6% vs 5.6%). MPV at 16℃ increased 43.2% compared with at 37℃, but had no changes along with the change of loading time and concentration. When loading at 37℃, MPV change was positively related with loading time and concentration, and loading time and loading concentration had a cooperative effect. MPV had no changes in 1～4hours loading when trehalose loading concentration was under 50mM. MPV increased with the increase of loading time and concentration when trehalose loading concentration was above 50mM. When trehalose loading concentration was 45mM, loading efficiency at 37℃ reached 13.86% 4 hours aftr loading. Conclusion The basic parameter of trehalose loading technique of human platelets was loading temperature at 37℃, loading time for 4 hours, and loading concentration of 45mM.