Development of a simplified platelet cryopreservation method: An in vitro investigation of reducing the DMSO concentration to allow administration without its pre-transfusion removal.

IF 1.8 4区医学 Q3 HEMATOLOGY

Vox Sanguinis Pub Date : 2025-03-01 Epub Date: 2025-01-06 DOI:10.1111/vox.13789

Lacey Johnson, Pearl Lei, Christopher Roan, Denese C Marks

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引用次数: 0

Abstract

Background and objectives: The most widely used method of platelet cryopreservation requires the addition of 5%-6% dimethylsulphoxide (DMSO), followed by its pre-freeze removal via centrifugation, to minimize toxicity. However, this adds complexity to the pre-freeze and post-thaw processing. Accordingly, the aim of this study was to simplify platelet cryopreservation by reducing the DMSO concentration and omitting the requirement for pre-transfusion removal.

Materials and methods: Apheresis platelets were cryopreserved at -80°C according to standard blood-banking methods using 5.5% DMSO, with centrifugation, pre-freeze removal of DMSO and reconstitution in plasma following thawing (standard). In parallel, doses of DMSO (0%, 1.5%, 3%, 5.5%) were tested without centrifugation and reconstitution (no-wash). In vitro platelet quality was assessed by flow cytometry, aggregation, viscoelastic testing (thromboelastography [TEG]) and clot retraction.

Results: Many in vitro platelet quality parameters showed DMSO dose dependency using the no-wash protocol (recovery, annexin-V, TEG maximum amplitude [MA]). Platelets frozen using the no-wash method with 3% DMSO showed a higher abundance of GPIbα (3% DMSO no-wash median fluorescence intensity [MFI]: 228 ± 16; standard MFI: 184 ± 16; p = 0.0016) and less degranulation (reduced P-selectin-positive platelets and concentration of supernatant P-selectin) than platelets frozen using the standard method. All functional properties measured were comparable to those of platelets frozen using the standard method.

Conclusion: This study shows that improvements in cryopreserved platelet quality parameters can be obtained by removing the centrifugation processes (standard vs. 5.5% DMSO no-wash). A reduction in DMSO to 3% supports quality parameters, and if shown to be clinically acceptable, this cryopreservation method could improve platelet accessibility, as it is simpler and cheaper than the standard method.

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一种简化的血小板冷冻保存方法的发展：降低DMSO浓度的体外研究，使其在输血前不被清除。

背景和目的：最广泛使用的血小板冷冻保存方法需要添加5%-6%的二甲基亚砜（DMSO），然后通过离心将其冷冻前去除，以减少毒性。然而，这增加了冷冻前和解冻后处理的复杂性。因此，本研究的目的是通过降低DMSO浓度和省略输血前去除的要求来简化血小板冷冻保存。材料与方法：单采血小板按照标准血库方法-80°C低温保存，使用5.5% DMSO，离心，冷冻前去除DMSO，解冻后血浆中重建（标准）。同时，在不离心和不清洗的情况下检测DMSO（0%、1.5%、3%、5.5%）的剂量。体外血小板质量通过流式细胞术、聚集、粘弹性测试（血栓弹性成像[TEG]）和凝块缩回来评估。结果：许多体外血小板质量参数在无洗方案下显示DMSO剂量依赖性（回收率、膜联蛋白v、TEG最大振幅[MA]）。用3% DMSO免洗法冷冻的血小板显示更高的GPIbα丰度(3% DMSO免洗中位荧光强度[MFI]: 228±16；标准MFI: 184±16；p = 0.0016)和较少的脱粒（p -选择素阳性血小板和p -选择素上清浓度降低）比使用标准方法冷冻的血小板。所有测量的功能特性都与使用标准方法冷冻的血小板相当。结论：本研究表明，通过去除离心工艺（标准与5.5% DMSO免洗）可以改善冷冻保存的血小板质量参数。DMSO降低至3%支持质量参数，如果临床证明是可接受的，这种低温保存方法可以提高血小板可及性，因为它比标准方法更简单，更便宜。

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来源期刊

Vox Sanguinis 医学-血液学

CiteScore

4.40

自引率

11.10%

发文量

156

审稿时长

6-12 weeks

期刊介绍： Vox Sanguinis reports on important, novel developments in transfusion medicine. Original papers, reviews and international fora are published on all aspects of blood transfusion and tissue transplantation, comprising five main sections: 1) Transfusion - Transmitted Disease and its Prevention: Identification and epidemiology of infectious agents transmissible by blood; Bacterial contamination of blood components; Donor recruitment and selection methods; Pathogen inactivation. 2) Blood Component Collection and Production: Blood collection methods and devices (including apheresis); Plasma fractionation techniques and plasma derivatives; Preparation of labile blood components; Inventory management; Hematopoietic progenitor cell collection and storage; Collection and storage of tissues; Quality management and good manufacturing practice; Automation and information technology. 3) Transfusion Medicine and New Therapies: Transfusion thresholds and audits; Haemovigilance; Clinical trials regarding appropriate haemotherapy; Non-infectious adverse affects of transfusion; Therapeutic apheresis; Support of transplant patients; Gene therapy and immunotherapy. 4) Immunohaematology and Immunogenetics: Autoimmunity in haematology; Alloimmunity of blood; Pre-transfusion testing; Immunodiagnostics; Immunobiology; Complement in immunohaematology; Blood typing reagents; Genetic markers of blood cells and serum proteins: polymorphisms and function; Genetic markers and disease; Parentage testing and forensic immunohaematology. 5) Cellular Therapy: Cell-based therapies; Stem cell sources; Stem cell processing and storage; Stem cell products; Stem cell plasticity; Regenerative medicine with cells; Cellular immunotherapy; Molecular therapy; Gene therapy.