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负载海藻糖的人体血小板在冻干后仍能存活。

Human platelets loaded with trehalose survive freeze-drying.

作者信息

Wolkers W F, Walker N J, Tablin F, Crowe J H

机构信息

Section of Molecular and Cellular Biology, University of California, Davis, CA 95616, USA.

出版信息

Cryobiology. 2001 Mar;42(2):79-87. doi: 10.1006/cryo.2001.2306.

Abstract

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 degrees C, 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 microg/ml), ADP (20 micromM), and ristocetin (1.6 mg/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.

摘要

根据联邦法规,人体血小板在血库中储存5天后即被丢弃。这种短寿命导致了血小板的长期短缺,这一问题在免疫抑制患者中尤为严重,比如艾滋病患者。我们在此报告,血小板可以通过用海藻糖进行冻干来保存,海藻糖是一种在天然耐干燥的生物体中高浓度存在的糖类。我们认为这些发现将消除血小板的储存问题。海藻糖在37摄氏度时能被人体血小板迅速摄取,摄取效率达到50%或更高。液相内吞作用在海藻糖的这种高效摄取中起重要作用,但也可能涉及其他机制。负载海藻糖的血小板成功进行了冻干,完整血小板的回收率很高。从气相复水后,存活率达到85%。这些血小板对凝血酶(1 U/ml)、胶原(2 μg/ml)、ADP(20 μM)和瑞斯托霉素(1.6 mg/ml)等激动剂的反应与新鲜对照血小板几乎相同。傅里叶变换红外光谱分析表明,负载海藻糖的血小板在冻干、预水合和复水后的膜和蛋白质成分与新鲜血小板非常相似。

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