[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.
Liu
Jing-Han
JH
Department of Blood Transfusion of General Hospital, Clinical Blood Trasfusion Centre of PLA, Beijing 100853, China. jinghan_liu@301hospital.cm.cn
Zhou
Jun
J
Ouyang
Xi-Lin
XL
Li
Xi-Jin
XJ
Lu
Fa-Qiang
FQ
D001792:Blood Platelets / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D001793:Blood Preservation / Q000379:methods*; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000378:metabolism / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005612:Freeze Drying; D006801:Humans; D014199:Trehalose / Q000378:metabolism / Q000494:pharmacology*
D003451:Cryoprotective Agents; D014199:Trehalose