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A comparative study of platelet storage lesion in platelet-rich plasma under cryopreservation

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Abstract

Platelet-rich plasma (PRP) has significant potential for various applications and holds clinical value in regenerative medicine. Cryopreservation is used to extend the preservation period of PRP, facilitating its clinical application. However, the potential negative effects of long-term cryopreservation on platelet storage lesion are still uncertain. In this study, PRP was stored at − 30 °C or − 80 °C. Platelet count, apoptosis, reactive oxygen species (ROS) content, and CD62P expression were assessed on the 14th and 28th days. The study also evaluated platelet mitochondria morphology and function, serotonin (5-HT) secretion by platelets, and the inflammatory activating effect of cryopreserved platelets in PRP. The results showed that there were no significant differences in platelet count, the content of 5-HT, and inflammatory effects between fresh PRP and PRP cryopreserved at both − 30 °C and − 80 °C. However, there was an increase in ROS level, apoptosis, and CD62P level after cryopreservation at both temperatures. Additionally, the levels of ROS, apoptosis, and CD62P in platelets were similar after storage at − 30 °C and − 80 °C. The main difference observed was that the morphology and function of mitochondria were severely damaged after storage at − 30 °C, while they were less affected at − 80 °C. Based on these findings, it can be concluded that storing PRP at − 80 °C is more suitable for achieving a better therapeutic effect in clinical applications, but cryopreservation could not replace the current standard.

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Acknowledgements

The authors would like to thank all blood volunteers for their participation in this study. We thank the Electron Microscopy Center of Fourth Military Medical University for this study.

Funding

The present work was supported by the National Natural Science Foundation of China (No. 82170226).

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  1. Department of Transfusion Medicine, Xijing Hospital, Fourth Military Medical University, Xi’an, Shaanxi, China

    E. Dang, Yutong Chen, Wenting Wang, Lingling Zhang, Ning An, Wen Yin, Jing Yi & Yaozhen Chen

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Contributions

ED, YTC, and WTW performed the experiments and analyzed and interpreted the data. YTC and LLZ drafted the manuscript. NA and WY contributed reagents. YZC and JY conceived and designed the experiments. YZC and YTC revised the manuscript. All authors read and approved the final version of the manuscript.

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Correspondence to Yaozhen Chen.

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All authors’ consent for the use of biospecimens was obtained in accordance with the Declaration of Helsinki. The experimental procedures were performed according to the requirements of the Ethics Committee of Xijing Hospital.

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Informed consent was obtained from all blood volunteers for being included in the study.

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The authors declare no competing interests.

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E. Dang, Yutong Chen, and Wenting Wang contributed equally as the first authors. Yaozhen Chen and Jing Yi contributed equally as the senior authors.

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277_2023_5580_MOESM1_ESM.tif

Supplementary file1 (TIF 8288 KB) Detection of the count of platelets in PRP under different storage temperature and storge time. (A) A schematic diagram of PRP treatment. (B) The count of platelets in PRP on 0 day, 14th day, 28th day at -30℃ (N = 5). (C) The count of platelets in PRP on 0 day, 14th day, 28th day at -80℃ (N = 5). (D) The count of platelets in PRP at -30℃ and -80℃ on 14th day (N = 5). (E) The count of platelets in PRP at -30℃ and -80℃ on 28th day (N = 5). ns, no statistical significance. PLT: platelets in PRP; 0d: platelets in fresh PRP, as control group; 14d: platelets in PRP were stored on the 14th day; 28d: platelets in PRP were stored on the 28th day; -30℃: platelets in PRP stored at -30℃; -80℃: platelets in PRP stored at -80℃.

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Dang, E., Chen, Y., Wang, W. et al. A comparative study of platelet storage lesion in platelet-rich plasma under cryopreservation. Ann Hematol 103, 631–643 (2024). https://doi.org/10.1007/s00277-023-05580-0

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