Preserving extracellular vesicles for biomedical applications: consideration of storage stability before and after isolation

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Abstract

Extracellular vesicles (EVs) are nanovesicles released by various cell types. EVs are known for cell-to-cell communications and have potent biological activities. Despite great progress in recent years for studies exploring the potentials of EVs for early disease detection, therapeutic application and drug delivery, determination of the favorable storage conditions of EVs has been challenging. The understanding of the impact of storage conditions on EVs before and after isolation is still limited. Storage may change the size, number, contents, functions, and behaviors of EVs. Here, we summarized current studies about the stability of EVs in different conditions, focusing on temperatures, durations, and freezing and thawing cycles. –80 °C seems to remain the most favorable condition for storage of biofluids and isolated EVs, while isolated EVs may be stored at 4 °C shortly. Lyophilization is promising for storage of EV products. Challenges remain in the understanding of storage-mediated change in EVs and in the development of advanced preservation techniques of EVs.

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Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines

Clotilde Théry, Kenneth W Witwer, Elena Aikawa … (2019)

ABSTRACT The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points.

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Communication by Extracellular Vesicles: Where We Are and Where We Need to Go.

Mercedes Tkach, Clotilde Théry, Beatriz Cara (2016)

In multicellular organisms, distant cells can exchange information by sending out signals composed of single molecules or, as increasingly exemplified in the literature, via complex packets stuffed with a selection of proteins, lipids, and nucleic acids, called extracellular vesicles (EVs; also known as exosomes and microvesicles, among other names). This Review covers some of the most striking functions described for EV secretion but also presents the limitations on our knowledge of their physiological roles. While there are initial indications that EV-mediated pathways operate in vivo, the actual nature of the EVs involved in these effects still needs to be clarified. Here, we focus on the context of tumor cells and their microenvironment, but similar results and challenges apply to all patho/physiological systems in which EV-mediated communication is proposed to take place.

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Bovine milk-derived exosomes for drug delivery.

Radha Munagala, Farrukh Aqil, Jeyaprakash Jeyabalan … (2016)

Exosomes are biological nanovesicles that are involved in cell-cell communication via the functionally-active cargo (such as miRNA, mRNA, DNA and proteins). Because of their nanosize, exosomes are explored as nanodevices for the development of new therapeutic applications. However, bulk, safe and cost-effective production of exosomes is not available. Here, we show that bovine milk can serve as a scalable source of exosomes that can act as a carrier for chemotherapeutic/chemopreventive agents. Drug-loaded exosomes showed significantly higher efficacy compared to free drug in cell culture studies and against lung tumor xenografts in vivo. Moreover, tumor targeting ligands such as folate increased cancer-cell targeting of the exosomes resulting in enhanced tumor reduction. Milk exosomes exhibited cross-species tolerance with no adverse immune and inflammatory response. Thus, we show the versatility of milk exosomes with respect to the cargo it can carry and ability to achieve tumor targetability. This is the first report to identify a biocompatible and cost-effective means of exosomes to enhance oral bioavailability, improve efficacy and safety of drugs.

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Author and article information

Journal

Journal ID (nlm-ta): Drug Deliv

Journal ID (iso-abbrev): Drug Deliv

Title: Drug Delivery

Publisher: Taylor & Francis

ISSN (Print): 1071-7544

ISSN (Electronic): 1521-0464

Publication date (Electronic, pub): 14 July 2021

Publication date (Electronic, collection): 2021

Volume: 28

Issue: 1

Pages: 1501-1509

Affiliations

[a ]Department of Clinical Medicine, Grade 2018, Xiangya School of Medicine of Central South University , Changsha, China

[b ]Clinical Nursing Teaching and Research Section, Second Xiangya Hospital of Central South University , Changsha, China

[c ]Hunan Testing Institute for Medical Devices , Changsha, China

Author notes

CONTACT Ya-Min Li aminny@ 123456csu.edu.cn Clinical Nursing Teaching and Research Section, Second Xiangya Hospital of Central South University , Changsha, Hunan 410011, China

Zhuhui Wang 867884013@ 123456qq.com Hunan Testing Institute for Medical Devices , 510 Tiyuan Road, Changsha, Hunan 410014, China

Article

Publisher ID: 1951896

DOI: 10.1080/10717544.2021.1951896

PMC ID: 8281093

PubMed ID: 34259095

SO-VID: 36b94eab-0135-494b-9d3b-3a18a6112629

License:

This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Preserving extracellular vesicles for biomedical applications: consideration of storage stability before and after isolation

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Most cited references 52

Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines

Communication by Extracellular Vesicles: Where We Are and Where We Need to Go.

Bovine milk-derived exosomes for drug delivery.

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