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用于治疗应用的人血小板浓缩物的生物加工,以生成裂解物和细胞外囊泡。

Bioprocessing of human platelet concentrates to generate lysates and extracellular vesicles for therapeutic applications.

作者信息

Yeh Wei-Ting, Yu Ezrin Yi-Ling, Lu Ya-Hsuan, Livkisa Dora, Burnouf Thierry, Lundy David J

机构信息

School of Biomedical Engineering, Taipei Medical University, 301 Yuantong Road, Taipei 235603, Taiwan.

International PhD Program in Biomedical Engineering, Taipei Medical University, 301 Yuantong Road, Taipei 235603, Taiwan.

出版信息

MethodsX. 2024 Jul 9;13:102822. doi: 10.1016/j.mex.2024.102822. eCollection 2024 Dec.

DOI:10.1016/j.mex.2024.102822
PMID:39105089
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11299553/
Abstract

This work describes protocols for preparing specific forms of human platelet lysates from pooled platelet concentrates (PCs) and the isolation of platelet-derived extracellular vesicles (p-EVs). Clinical-grade PCs can be sourced from blood establishments immediately following expiration for transfusion use. Here, we describe methods to process PCs into specific lysates from which p-EVs can be isolated. Each lysate type is prepared using platelet activation and processing methods which produce distinct products that may be useful in different applications. For example, serum-converted platelet lysate (SCPL)-EVs were recently shown to have powerful therapeutic properties following myocardial infarction in mice. EVs can be isolated from all products using size exclusion chromatography, producing pure and consistent p-EVs from multiple batches. Together, these methods allow isolation of p-EVs with excellent potential for clinical and preclinical applications.•Platelet concentrates (PCs) obtained from local blood establishments are reliable and sustainable sources to generate biomaterials.•We outline five distinct methods of platelet lysate generation and one method for extracellular vesicle isolation.•Each platelet lysate form has different biological properties which may be suitable for certain applications.

摘要

本研究描述了从混合血小板浓缩物(PCs)制备特定形式的人血小板裂解物以及分离血小板衍生细胞外囊泡(p-EVs)的方案。临床级PCs可在输血用有效期刚过之后从血液机构获取。在此,我们描述了将PCs加工成可从中分离p-EVs的特定裂解物的方法。每种裂解物类型都是使用血小板激活和加工方法制备的,这些方法会产生不同的产物,可能在不同应用中有用。例如,血清转化血小板裂解物(SCPL)-EVs最近被证明在小鼠心肌梗死后具有强大的治疗特性。使用尺寸排阻色谱法可从所有产物中分离出EVs,从而从多个批次中产生纯净且一致的p-EVs。总之,这些方法能够分离出在临床和临床前应用中具有巨大潜力的p-EVs。

•从当地血液机构获得的血小板浓缩物(PCs)是生成生物材料的可靠且可持续的来源。

•我们概述了五种不同的血小板裂解物生成方法和一种细胞外囊泡分离方法。

•每种血小板裂解物形式都具有不同的生物学特性,可能适用于某些应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b522/11299553/1683e1680308/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b522/11299553/4a2700c5bd7c/ga1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b522/11299553/b5392a9a5b5e/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b522/11299553/9d79d6a7d275/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b522/11299553/1683e1680308/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b522/11299553/4a2700c5bd7c/ga1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b522/11299553/b5392a9a5b5e/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b522/11299553/9d79d6a7d275/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b522/11299553/1683e1680308/gr3.jpg

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Optimization of extracellular vesicle isolation and their separation from lipoproteins by size exclusion chromatography.通过尺寸排阻色谱法优化细胞外囊泡的分离及其与脂蛋白的分离。
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Calcium chloride declotted human platelet lysate promotes the expansion of mesenchymal stromal cells and allows manufacturing of immunomodulatory active extracellular vesicle products.
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Life (Basel). 2024 Dec 27;15(1):12. doi: 10.3390/life15010012.
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