Current status of extracellular vesicle technology in industrialization and commercialization

Abstract

Extracellular vesicles (EVs) are nanovesicles secreted by most cells types that play essential roles in intercellular communication and molecular exchange. Due to their diverse cellular origins, small molecule size, high stability, excellent biocompatibility, and cell targeting, EVs have emerged as promising tools for disease diagnosis and therapy, targeted drug delivery, and regenerative medicine. This review provides an overview of the biochemical characteristics, production methods, and industrial development of EVs based on patent filings and registered clinical trials related to EVs from 2015 to 2024. We further analyze major translational bottlenecks hindering the commercialization of EVs, emphasizing challenges related to batch-to-batch reproducibility, methodological standardization, manufacturing scalability, and regulatory pathway clarity. Finally, we highlight new engineering approaches including 3D hollow-fiber bioreactors, microfluidic isolation platforms, and genetic bioengineering that hold significant potential to address current limitations in EV manufacturing and clinical translation.

Supplementary Information

The online version contains supplementary material available at https://doi.org/10.1186/s11671-026-04862-6.

Highlights

To summarize the composition, functions, and applications of EVs.

To analyze EV-related patents and clinical trials from the last ten years.

To provide an overview of the EV market and manufacturing sectors.

To explore the translational challenges and commercial barriers of EV-based technology.

Supplementary Information

The online version contains supplementary material available at https://doi.org/10.1186/s11671-026-04862-6.

Introduction

Extracellular vesicles (EVs) are a heterogeneous group of particles released from cells, enclosed by a lipid bilayer, and incapable of replication on their own. According to the latest MISEV2023 guidelines published by the International Society for Extracellular Vesicles (ISEV) [1], EVs are currently classified into two main subtypes based on their origin or size. Exosomes, ranging from 30 to 150 nm in diameter, are derived from the internal compartments of the cell, while microvesicles, range from 100 to 1000 nm in diameter, are released by the outward budding of the plasma membrane [2]. Additionally, several specialized terms are used to describe EVs that are formed during specific cellular processes, such as programmed cell death (referred to as ‘apoptotic bodies’) [1]. Because it is difficult to completely distinguish between these types of small vesicles, all such vesicles are referred to as EVs in this review.

Owing to their unique structure, EVs offer significant potential for clinical applications. The number of registered patents and ongoing clinical trials involving EV-derived products has increased. However, the widespread clinical and industrial use of EVs is still limited by major obstacles, especially their molecular and structural diversity. Robust, scalable, and reproducible purification methods for EV-related products require further refinement and standardization to ensure the quality and safety. Therefore, a comprehensive assessment of the current status in industrialization and commercialization of EV-based technology is essential.

Characteristics and function of EVs

EVs were first described as membrane vesicles released from sheep reticulocytes by Pan and Johnstone in 1983 [3]. They encapsulate various components, including DNA, RNA, lipids, and proteins, depending on their cellular origin [4] (Fig. 1A). To date, the ExoCarta database (http://www.exocarta.org) has cataloged 13,476 proteins, 3,408 messenger RNAs (mRNAs), 10,755 miRNAs, and 3,846 lipids within EVs. Although different cell types secrete distinct proteins into EVs, protein conservation among various EV types is as high as 80% Membrane proteins such as CD9, CD63, and CD81, along with cytosolic proteins including tumor susceptibility gene 101 (TSG101) and apoptosis-linked gene 2-interacting protein X (Alix), serve as biomarkers for EV characterization [5–6]. EVs are rich in nucleic acids, including noncoding RNAs, mRNAs and mitochondrial DNA [7], most of which consist of approximately 200-base pair degraded fragments [8].

Fig. 1.

Composition and main clinical applications of extracellular vesicles (EVs)

EVs may mediate specific cell to cell communication and participate in numerous biological processes [4, 9–10]. Due to their relatively small size, EVs can transport biological molecules across the blood–brain barrier, making them promising diagnostic markers and therapeutic agents for brain-related diseases [11, 12]. Clinically, EVs are primarily utilized as biomarkers, cell-free therapeutic agents, drug-delivery vehicles, and cancer vaccines [13–16] (Fig. 1B). The diagnostic applications of EVs is rapidly expanding to diseases involving the central nervous system (CNS) [17], liver [18], kidneys [19], and lungs [20]. EVs can deliver drugs to target cells for various purposes, such as enhancing cell sensitivity and efficiency [21]. Because EVs are rich in tumor antigens, including major histocompatibility complex (MHC) class I and II molecules, they are excellent candidates for cell-free cancer vaccines [22]. EVs derived from stem cells hold significant potential for therapeutic applications in tissue regeneration [23].

Preparation and Identification of EVs

Several techniques, including ultracentrifugation(UC), size-based isolation, polymer precipitation, and immunoaffinity capture, have been used to isolate and purify EVs [24]. Among these, UC remains the most commonly used isolation method and is widely regarded as the gold standard for EV extraction [25]. This technique separates components based on differences in size and density, making it particularly suitable for processing large-volume samples [26]. However, the limitations of UC, including its time-intensive procedures, high cost, potential structural damage to EVs, particle aggregation, and co-isolation of lipoproteins, influence both EVs yield and purity [27–29]. Microfluidics offers an alternative approach for isolating EVs. The major advantage of microfluidics is that it allows for both isolation and characterization to be carried out on a single chip, enabling the isolation of EVs from minimal sample volume and the modulation of specific parameters [30–31]. Nevertheless, current microfluidic approaches still face challenges in achieving complete separation of EVs from lipoprotein contaminants [32]. Furthermore, fabrication of microfluidic chips remains technically demanding, time-consuming, and reliant on specialized expertise.

EVs are characterized by evaluating their physical properties and analyzing the composition of their biomolecular constituents [33]. Physical characterization techniques, including nanoparticle tracking analysis (NTA), dynamic light scattering (DLS), electron microscopy, and tunable resistive pulse sensing, are used to determine the particle size distribution and concentration. Biochemical identification of EVs is commonly performed via fluorescence labeling, immunoblot, and high-resolution proteomic profiling [34–36]. Conventional protein analysis methods, such as Western blotting and enzyme-linked immunosorbent assays (ELISAs), typically demand large sample volumes, involve multistep processing, and require specialized instruments. Therefore, their implementation in clinical settings remains challenging, particularly in large-scale patient cohorts studies or the quantification of low-abundance molecular markers. To address these technical limitations, next-generations biosensors are being developed. Unlike conventional protein detection methods, these biosensors utilize distinct sensing mechanisms and are designed to identify a broad spectrum of EVs that differ in size and molecular composition [37].

Analysis of EVs-related patents and clinical trials

The number of patents related to EVs has increased rapidly over the past decade, highlighting key research areas and the significant application potential in this field. A total of 10,743 patents related to EVs were registered in the PatentScope database (https://patentscope.wipo.int) between 2015 and 2024 using the keyword “exosome* or extracellular vesicle*”, with overlapping patents excluded. The data in Fig. 2A suggest a growing trend in the number of patent publications, reaching a maximum of 1,889 in 2024. Geographically, EV-related patents are predominantly concentrated in China (30.9%), the United States (16.1%), Europe (9.4%), and the Republic of Korea (5.4%), with 3,323, 1,735, 1,014, and 576 patents, respectively (Fig. 2B). This distribution reflects not merely regional technological advantages but rather divergent national strategic priorities in nanomedicine, and differential patenting landscapes. By technical domain, patents are distributed across five categories: therapeutics (34.3%), purification methods (22.0%), diagnostics (10.4%), drug delivery (4.7%), and vaccines (3.7%) (Fig. 2C). Notably, the majority of patents are concentrated in therapeutic applications and purification methods, particularly in extraction and purification techniques, protein engineering modifications, and carrier-based drug loading. These efforts aim to address challenges related to downstream processing technical obstacles, rather than directly focusing on the final drugs. This highlights the industry’s strategic focus on overcoming downstream processing bottlenecks, which remain a major obstacle to clinical translation.

Fig. 2.

EV-derived patents between 2015–2024. A Number of EV-derived patents. B Origin countries of EV-derived patents. C Field distribution ratios of EV-derived patents. D Phases of clinical trial stage

The global clinical trial databases from 2015 to 2024 were screened for EV-derived products by searching clinicaltrials.gov (https://clinicaltrials.gov) using the keywords “exosome” or “extracellular vesicle”. A total of 322 registered clinical trials were identified after removing duplicates (Supplemental Table 1). Among these, China ranks first globally, with 85 trials accounting for 26.4%, followed closely by the United States with 65 trials (20.2%). Italy leads among European countries, conducting 31 trials, followed by France with 20 trials (6.2%). A detailed analysis of the trial phases reveals a significant translational bottleneck. A total of 166 intervention trials were conducted, comprising Early Phase I (5.4%, 9), Phase I (17.5%, 29), Phase I/Phase II (16.3%, 27), Phase II (7.2%, 12 ), Phase II/Phase III (1.8%, 3), Phase III (1.2%, 2), and Phase IV (1.2%, 2) trials (Fig. 2D). More than 40% of intervention trials remain in the early phases (Phase I, Phase I/II, and Phase II), primarily focusing on verifying safety and tolerability. This indicates that although EVs demonstrate acceptable safety profiles in early small-scale cohorts, progression to Phase III trials or commercialization is significantly hindered by several challenges. As bioactive substances, EVs have complex compositions and unclear mechanisms of action, which increase the uncertainty of clinical trials. Additionally, due to their molecular heterogeneity and low secretion levels from cells, it is difficult to ensure batch-to-batch consistency and achieve high-purity preparations. These factors make it challenging to meet Good Manufacturing Practice (GMP) standards and impede the development of large-scale clinical trials.

Market overview and manufacturing industries of EVs

The rising global burden of chronic diseases, such as cancer, cardiovascular diseases, and neurological disorders, along with the growing need for screening and diagnosis, are key factors driving the demand in the global EV diagnostic and therapeutic market in the coming years. Data from marketresearchfuture.com (https://www.marketresearchfuture.com) indicates that the exosome therapeutic market was valued at USD 1.26 billion in 2024 and is projected to grow from USD 1.39 billion in 2025 and USD 3.71 billion by 2035, representing a compound annual growth rate( CAGR) of 10.29% between 2025 and 2035.

Within the EV therapeutics market, different product types display distinct market share distributions. Diagnostic products currently dominate the market, driven by increasing demand for early disease detection and advancements in precision medicine. In terms of growth trends, demand for therapeutic products is emerging as a significant driver within the EV therapeutics segment (Fig. 3A). According to the “Exosome Therapeutics Market Report” (https://www.marketresearchfuture.com), the revenue generated from exosome applications in cancer treatment is expected to increase from USD 0.38 billion in 2025 to USD 1.00 billion by 2035, reflecting strong market demand for innovative cancer therapies utilizing EVs. Similarly, the application of EVs in neurological disorders is projected to grow from a market value of USD 0.19 billion in 2025 to USD 0.57 billion by 2035 (Fig. 3B).

Fig. 3.

An overview of the exosome therapeutic market size in 2025 and 2035 (unit: one billion US dollars). A Exosome therapeutics market of diagnostic and therapeutic products. B Exosome therapeutics market of cancer treatment and neurological disorders in 2025 and 2035

Currently, no EV drugs are available on the market. However, CAP-1002 (Capricor) has completed phase II clinical trials for patients with Duchenne muscular dystrophy, and its developers are preparing to submit a Biologics License Application (BLA), indicating that major breakthroughs in EV therapeutics are imminent. CAP-1002 consists of cardiosphere-derived cells, which modulate immune system activity and promote cell regeneration [38]. ExoFlo is an EV product derived from human bone marrow mesenchymal stem cells (MSCs). Direct Biologics is currently conducting a global phase III EXTINGUISH ARDS clinical trial to evaluate the efficacy of ExoFlo treatment in hospitalized adults with moderate to severe acute respiratory distress syndrome (ARDS). Additionally, the company has initiated early-phase clinical trials of ExoFlo for treating ulcerative colitis and Crohn’s disease and has extended clinical trials for patients undergoing abdominal organ transplantation and those with severe ARDS. Intravenous delivery of ExoFlo appears to be safe and alleviates clinical symptoms in patients with complex regional pain syndrome (CRPS) [39].

Biotechnology and pharmaceutical companies hold a dominant position in the EV diagnostic and therapeutic market. Equipped with advanced technologies, these companies are actively promoting the clinical and commercial translation of EVs. The United States leads the global market, with key players such as Direct Biologics and Codiak BioSciences, Inc. (see Table 1). Codiak BioSciences was the first pharmaceutical company to conduct EV research. It completed phase I clinical trials of exoIL-12 and phase I/II clinical trials of exoSTING, achieving promising results in terms of safety and efficacy. However, the company shifted its focus to developing a coronavirus disease 2019 (COVID-19) vaccine and discontinued the development of exoSTING and exoIL-12. Unexpectedly, Codiak BioSciences declared bankruptcy and entered reorganization in 2023, likely due not to technical failure but to the impacts of the COVID-19 pandemic, clinical delay, and financing challenges. As the first publicly listed company in the EV field, Codiak BioSciences’ bankruptcy announcement has negatively affected the entire EV industry.

Table 1.

Key companies in the EV therapeutic market

Company	Drug	Cell (origin)	Disease	Trial phase	Country
Direct Biologics	ExoFIo	Human bone marrow MSCs	Moderate to severe ARDS	3	United States
Capricor Therapeutics	CAP-1002	Cardiosphere-derived cells	Duchenne muscular dystrophy	2	United States
VITTI Labs	VL-PX10, VL-P22	Umbilical cord MSCs	Coronavirus disease 2019 (COVID-19)-related ARDS	2	United States
Nano24med	EXO-CD24	Engineered EVs containing CD24	Mild-moderate ARDS	2	Israel
RION	PEP (Purified Exosome Product)/TISSEEL	Human platelets	Diabetic foot ulcers	2	Japan
ZEO ScientifiX	Zofin	Human amniotic fluid (HAF)	Chronic obstructive pulmonary disease COVID-19	1/2	United States
Aegle Therapeutics	AGLE-102	MSCs	Dystrophic epidermolysis bullosa	1/2	United States
Exo biologics	EXOB-001	MSCs	Premature infants with bronchopulmonary dysplasia	1/2	Belgium
PureTech	LYT-300	Milk	Acute anxiety disorder	1/2	United States
Codiak BioSciences	ExoSTING (CDK-002)	Normal fibroblasts	Advanced solid tumors	1/2	United States
	exoASO-STAT6 (CDK-004) [42–43]	Normal fibroblasts	Advanced hepatocellular carcinoma Gastric cancer metastatic to liver Colorectal cancer metastatic to liver	1
ILIAS Biologics	ILB-202	Engineered exosomes loaded with superrepressor IκBα	Acute kidney injury related to cardiac surgery	1	South Korea
Exopharm	PLEXOVAL II	human platelets	wound healing	1	Australia

Translational challenges and commercial bottlenecks

Although the number of patents related to EVs has increased rapidly over the past decade, the number of clinical trials involving EVs is growing slowly. Patents often highlight research hotspots and application potential. However, the transition of EV technologies from the laboratory to clinical application faces multifaceted translational challenges. To overcome these obstacles, systematic improvements need to be made in manufacturing processes and regulatory frameworks.

Quality control (QC) and the standardization of EV preparations are critical bottlenecks hindering widespread clinical adoption. QC standards for EVs are complex and require comprehensive evaluation and control of factors such as cell lines, culture conditions, stability, and controlled production processes to ensure quality and safety for therapeutic use. Due to their inherent biological heterogeneity, standardizing the EV production process to achieve consistent quality remains exceptionally challenging. As therapeutic carriers, EVs require validated, reproducible drug-loading capacities and stable in vivo release profiles. Current practices often exhibit high inter-batch variability. The guidelines in MISEV2023 are essential for establishing minimum quality control parameters, but the industry still lacks universally standardized operating procedures (SOPs) for EV isolation and characterization.

Achieving clinical-grade scalability requires transitioning from conventional small-scale methods to integrated, high-yield systems. UC remains the gold standard, but its prolonged processing time and high shear forces often cause vesicle damage, particle aggregation, and co-separation of lipoproteins. These limitations make UC unsuitable for commercial scalability [42–43]. To overcome these challenges, the field must adopt advanced manufacturing approaches. Novel three-dimensional (3D) cell culture technologies, particularly hollow-fiber 3D bioreactors, have demonstrated superior capabilities in producing high concentrations of EVs without altering cellular morphology [44–48]. Furthermore, combining isolation techniques, such as ligand-based exosome affinity purification (LEAP) chromatography, presents a promising strategy to isolate clinical-grade EVs at an industrial scale while preserving structural integrity [49]. Additionally, various engineering approaches have been explored for the surface modification of EVs using specific targeted molecules to enhance their targeting specificity and prevent clearance by the immune system [50–51] .

As a frontier biomedical technology, EV therapeutics face an evolving and increasingly stringent regulatory environment. Regulatory frameworks worldwide require rigorous adherence to GMP standards, originally developed for the pharmaceutical and food industries, to minimize contamination and ensure continuous compliance. Recently, regulatory bodies such as the U.S. Food and Drug Administration (FDA) have imposed stricter criteria for EV products, demanding comprehensive clinical data and robust evidence of manufacturing stability. For example, the FDA’s rejection of the BLA for CAP-1002 highlights the critical necessity for products to meet stringent biosimilarity and quality consistency standards. Moving forward, the successful commercialization of EVs will depend heavily on biotechnology firms proactively aligning with these dynamically tightening regulatory expectations.

Conclusion

EVs represent a promising frontier in nanomedicine and precision medicine due to their unique structure. While they serve as potential biomarkers and effective drug delivery agents, the clinical translation of EV products is currently undergoing a critical transition from conceptual exploration to early-stage commercialization. Optimism regarding their economic potential must be balanced against significant industrial challenges. Recent advancements in 3D cultivation, microfluidics, and engineered targeting offer encouraging progress. However, the definitive realization of EV therapies depends on establishing universally accepted QC standards and GMP-compliant, scalable manufacturing processes.

Supplementary Information

Below is the link to the electronic supplementary material.

Author contributions

Yuhao Wu contributed manuscript text .

Funding

Not applicable.

Data availability

No datasets were generated or analysed during the current study.

Declarations

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Consent to publish

Its publication has been approved by all co-authors, and the responsible authorities at the institute where the work has been carried out.

Dual publication

This manuscript has not been published or presented elsewhere in part or in its entirety and is not under consideration by another journal.

Competing interests

The authors declare no competing interests.