Human platelets loaded with trehalose survive freeze-drying

被引:328
作者
Wolkers, WF
Walker, NJ
Tablin, F
Crowe, JH
机构
[1] Univ Calif Davis, Sch Vet Med, Sect Mol & Cellular Biol, Davis, CA 95616 USA
[2] Univ Calif Davis, Sch Vet Med, Sect Anat Physiol & Cell Biol, Davis, CA 95616 USA
关键词
desiccation-tolerance; freeze-drying; glasses; platelets; trehalose;
D O I
10.1006/cryo.2001.2306
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Human blood platelets are stored in blood banks for 5 days, after which they are discarded, by Federal regulation. This short lifetime has led to a chronic shortage of platelets, a problem that is particularly acute in immunosuppressed patients, such as those with AIDS. We report here that platelets can be preserved by freeze-drying them with trehalose, a sugar Found at high concentrations in organisms that naturally survive drying. We suggest that these findings will obviate the storage problem with platelets. Trehalose is rapidly taken up by human platelets at 37 degreesC, with loading efficiencies of 50% or greater. Fluid-phase endocytosis plays an important role in this efficient uptake of trehalose, but other mechanisms may also be involved. Trehalose-loaded platelets were successfully freeze-dried, with excellent recovery of intact platelets. Rehydration From the vapor phase led to a survival rate of 85%. The response of these platelets to the agonists thrombin (1 U/ml), collagen (2 mug/ml), ADP (20 muM), and ristocetin (1.6 ms/ml) was almost identical to that of fresh, control platelets. Analysis by Fourier transform infrared spectroscopy demonstrated that the membrane and protein components of trehalose-loaded platelets after freeze-drying, prehydration, and rehydration were remarkably similar to those of fresh platelets. (C) 2001 Academic Press.
引用
收藏
页码:79 / 87
页数:9
相关论文
共 21 条
[1]   Trehalose: A cryoprotectant that enhances recovery and preserves function of human pancreatic islets after long-term storage [J].
Beattie, GM ;
Crowe, JH ;
Lopez, AD ;
Cirulli, V ;
Ricordi, C ;
Hayek, A .
DIABETES, 1997, 46 (03) :519-523
[2]   Cryopreservation of human platelets with dimethyl sulfoxide: Changes in biochemistry and cell function [J].
Bock, M ;
Schleuning, M ;
Heim, MU ;
Mempel, W .
TRANSFUSION, 1995, 35 (11) :921-924
[3]  
CROWE J H, 1989, P1
[4]   Are lipid phase transitions responsible for chilling damage in human platelets? [J].
Crowe, JH ;
Tablin, F ;
Tsvetkova, N ;
Oliver, AE ;
Walker, N ;
Crowe, LM .
CRYOBIOLOGY, 1999, 38 (03) :180-191
[5]   The role of vitrification in anhydrobiosis [J].
Crowe, JH ;
Carpenter, JF ;
Crowe, LM .
ANNUAL REVIEW OF PHYSIOLOGY, 1998, 60 :73-103
[6]  
CROWE JH, 1992, ANNU REV PHYSIOL, V54, P579, DOI 10.1146/annurev.ph.54.030192.003051
[7]   Intracellular trehalose improves the survival of cryopreserved mammalian cells [J].
Eroglu, A ;
Russo, MJ ;
Bieganski, R ;
Fowler, A ;
Cheley, S ;
Bayley, H ;
Toner, M .
NATURE BIOTECHNOLOGY, 2000, 18 (02) :163-167
[8]   GEOMETRY OF NORMAL MAMMALIAN PLATELETS BY QUANTITATIVE MICROSCOPIC STUDIES [J].
FROJMOVIC, MM ;
PANJWANI, R .
BIOPHYSICAL JOURNAL, 1976, 16 (09) :1071-1089
[9]   Trehalose expression confers desiccation tolerance on human cells [J].
Guo, N ;
Puhlev, I ;
Brown, DR ;
Mansbridge, J ;
Levine, F .
NATURE BIOTECHNOLOGY, 2000, 18 (02) :168-171
[10]   DOES FOURIER-TRANSFORM INFRARED-SPECTROSCOPY PROVIDE USEFUL INFORMATION ON PROTEIN STRUCTURES [J].
HARIS, PI ;
CHAPMAN, D .
TRENDS IN BIOCHEMICAL SCIENCES, 1992, 17 (09) :328-333