Loading Trehalose into Platelets by Ultrasound

Abstract: To investigate the influence of ultrasound on loading trehalose into human platelets， human platelets at concentration of about 1×10⁹ cells/ml were hatched in solution that contained 50 mmol/L trehalose， 100 mmol/L NaCl， 10 mmol/L KCl， 10 mmol/L EGTA and 10 mmol/L imidazole. The hatching lasted for 4 h at 37 ℃. 800 kHz ultrasound with several intensities was added on the samples for varying periods. The platelet sample hatched without ultrasound was chosen as the reference. The quantities of sugar were measured through vitriol-authrone method with spectrophotometer and used to calculate trehalose concentration that had been loaded into platelets. Results demonstrated that when the ultrasound intensity I=0.8 W/cm² and the radiation duration was one hour， the trehalose concentration loaded into platelets was the highest， (17.40±.90) mmol/L. This value was 54.3% higher than the reference group， (11.27±2.53) mmol/L. Through microscope observation， it was found that the platelets after loading trehalose with ultrasound kept integrity and normal shape， and showed almost no difference with fresh platelets. Appropriate ultrasound radiation can effectively enhance the loading of trehalose into human platelets， and fundamental conditions for further research on freeze-drying of platelets.
    

CSTR: 32200.14.cjcb.2008.01.0024