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新型冷冻人血小板的制备方法及质量评价

[Preparation Method and Quality Evaluation of Novel Frozen Human Platelets].

作者信息

Zheng Yi-Zhe, Li Dong-Dong, Yan Geng-Wei, Wang Bao-Jian, Wang Ke, Wang Lei, Yan Shao-Duo, Li Yan-Hong, Fu Qiu-Xia, Sun Zhen-Wei

机构信息

988th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army, Zhengzhou 450042, Henan Province, China.

Institute of Health Services and Blood, Academy of Military Medical Sciences, Beijing 100039, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2024 Aug;32(4):1264-1270. doi: 10.19746/j.cnki.issn.1009-2137.2024.04.044.

Abstract

OBJECTIVE

To optimize the technical parameters related to the preparation of novel frozen human platelets and formulate corresponding protocol for its preparation.

METHODS

Novel frozen human platelets were prepared with O-type bagged platelet-rich plasma (PRP), the key technical parameters (DMSO addition, incubation time, centrifugation conditions, etc.) of the preparation process were optimized, and the quality of the frozen platelets was evaluated by routine blood tests, apoptosis rate, platelet activation rate and surface protein expression level.

RESULTS

In the preparation protocol of novel frozen human platelets, the operation of centrifugation to remove supernatant was adjusted to before the procedure of platelets freezing, and the effect of centrifugation on platelets was minimal when the centrifugation condition was 800× for 8 min. In addition, platelets incubated with DMSO for 30 min before centrifugation exhibited better quality after freezing and thawing. The indexes of novel frozen human platelets prepared with this protocol remained stable after long-term cryopreservation.

CONCLUSION

The preparation technique of novel frozen human platelets was established and the protocol was formulated. It was also confirmed that the quality of frozen platelets could be improved by incubating platelets with DMSO for 30 min and then centrifuging them at 800× for 8 min in the preparation of novel frozen human platelets.

摘要

目的

优化新型冷冻人血小板制备的相关技术参数,并制定相应的制备方案。

方法

采用O型袋装富血小板血浆(PRP)制备新型冷冻人血小板,对制备过程中的关键技术参数(二甲基亚砜添加量、孵育时间、离心条件等)进行优化,通过血常规、凋亡率、血小板活化率及表面蛋白表达水平对冷冻血小板质量进行评价。

结果

在新型冷冻人血小板制备方案中,将去除上清的离心操作调整至血小板冷冻步骤之前,离心条件为800×8分钟时离心对血小板的影响最小。此外,离心前用二甲基亚砜孵育血小板30分钟,冻融后血小板质量更佳。按此方案制备的新型冷冻人血小板各项指标经长期冻存后保持稳定。

结论

建立了新型冷冻人血小板的制备技术并制定了方案。同时证实,在新型冷冻人血小板制备过程中,血小板用二甲基亚砜孵育30分钟后,再以800×离心8分钟,可提高冷冻血小板质量。

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