冷冻血小板的生存能力和功能评估。

Evaluation of survival and functionality in frozen platelets.

作者信息

Rueda Jhon Alexander Avila, Acosta Jose, Rabinovich Oscar, Ceresetto José, Ernst Glenda, Duboscq Cristina

机构信息

Hospital Británico de Buenos Aires; Buenos Aires, Argentina; Hospital Privado Santa Clara De Asís, Salta, Argentina.

Hospital Británico de Buenos Aires; Buenos Aires, Argentina.

出版信息

Hematol Transfus Cell Ther. 2024 Apr-Jun;46(2):119-124. doi: 10.1016/j.htct.2023.01.002. Epub 2023 Jan 25.

DOI:10.1016/j.htct.2023.01.002

PMID:36717327

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11150498/

Abstract

OBJECTIVE

Prolong platelet survival and functionality up to 28 days.

METHODS

A sample of apheresis platelets was evaluated, distributed in 3 groups according to the cryopreservative solution used: DMSO5%+2%albumin; DMSO5%+NaCl0,9% and DMSO5%+Dextrose2%. They were then frozen at -80 °C and thawed at 7, 14 and 28 days. The in vitro survival and viability were assessed by the post-thaw platelet count and the CD41, CD61 and CD42a staining percentages by flow cytometry. The functionality was determined with the percentage of post-stimulation aggregation with 1Nm-thrombin using the Chromo-Log490 aggregometer. The control group (CG) consisted of fresh platelets under constant agitation at 22 °C.

RESULTS

A total of 72 platelet aliquots was analyzed. The CG presented a platelet-count of 1934 ± 0.5 × 10/L and a 100% viability. The percentages of CD41, CD61 and CD42a labeling were 99, 98.5 and 96.5%, respectively. The percentage of aggregation was 99%. On day 7 of the post-freezing, the platelet count for groups 1, 2 and 3 was 1,844 ± 102, 1,856 ± 76 and 1,752 ± 226, with the viability of 98, 96 and 95%, respectively. On day 14, the counts were 1,722 ± 238, 1,649 ± 215 and 1,578 ± 223 with the viability of 96, 95 and 94% and, on day 28, they were 1,602 ± 374, 1,438.6 ± 429 and 1,406.6 ± 436, with the viability of 96, 94 and 93%, respectively. Group1 presented a higher expression of membrane antigens. Aggregation percentages were 90, 98 and 89% at day 7, 88%, 98 and 87% at day 14 and 84%, 95 and 82% at day of the 28 post-freezing, respectively, with group2 presenting the best results.

CONCLUSION

The results support cryopreservation as a reasonable method to prolong platelet survival up to 28 days, maintaining its functionality and viability greater than 50%.

摘要

目的

将血小板的存活时间和功能延长至28天。

方法

对一份单采血小板样本进行评估，根据所使用的冷冻保存溶液分为3组：5%二甲基亚砜+2%白蛋白；5%二甲基亚砜+0.9%氯化钠；5%二甲基亚砜+2%葡萄糖。然后将它们在-80°C下冷冻，并在第7天、14天和28天解冻。通过解冻后血小板计数以及流式细胞术检测CD41、CD61和CD42a染色百分比来评估体外存活和活力。使用Chromo-Log490血小板聚集仪通过1N/m凝血酶刺激后聚集的百分比来测定功能。对照组（CG）由在22°C持续搅拌下的新鲜血小板组成。

结果

共分析了72份血小板样本。对照组的血小板计数为1934±0.5×10⁹/L，活力为100%。CD41、CD61和CD42a标记的百分比分别为99%、98.5%和96.5%。聚集百分比为99%。在冷冻后的第7天，第1组、第2组和第3组的血小板计数分别为1844±102、1856±76和1752±226，活力分别为98%、96%和95%。在第14天，计数分别为1722±238、1649±215和1578±223，活力分别为96%、95%和94%，在第28天，计数分别为1602±374、1438.6±429和1406.6±436，活力分别为96%、94%和93%。第1组的膜抗原表达较高。在冷冻后第7天、14天和28天的聚集百分比分别为90%、98%和89%，88%、98%和87%，84%、95%和82%，第2组的结果最佳。