Impact of storage conditions and duration on function of native and cargo-loaded mesenchymal stromal cell extracellular vesicles

Levy, Daniel; Jeyaram, Anjana; Born, Louis J.; Chang, Kyungho; Abadchi, Sanaz Nourmohammadi; Hsu, An; Solomon, Talia; Aranda, Amaya; Stewart, Stanford J.; He, Xiaoming; Harmon, John W.; Jay, Steven M.

doi:10.1016/j.jcyt.2022.11.006

Cited by 15 publications

(11 citation statements)

References 36 publications

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“…Extracellular vesicles (EVs), specifically those from mesenchymal stem cells (MSCs), have emerged as an intriguing therapeutic alternative to whole‐cell therapies in a multitude of applications (e.g., sepsis, cancer, wound healing) with additional promise as efficient drug delivery vehicles. [ 1 ] With an improved safety profile, [ 2 ] favorable storage requirements, [ 3 ] and the ability to traverse biological barriers, [ 4 ] MSC EVs represent a promising and effective alternative to their parental cells. However, there exist fundamental obstacles that hinder the translation of MSC EVs, including a lack of a rationally designed production platform and low therapeutic potency (i.e., low levels of endogenous EV cargos).…”

Section: Introductionmentioning

confidence: 99%

Mesenchymal Stem Cell Culture within Perfusion Bioreactors Incorporating 3D‐Printed Scaffolds Enables Improved Extracellular Vesicle Yield with Preserved Bioactivity

Kronstadt

Patel

Born

et al. 2023

Adv Healthcare Materials

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Extracellular vesicles (EVs) are implicated as promising therapeutics and drug delivery vehicles in various diseases. However, successful clinical translation will depend on the development of scalable biomanufacturing approaches, especially due to the documented low levels of intrinsic EV‐associated cargo that may necessitate repeated doses to achieve clinical benefit in certain applications. Thus, here the effects of a 3D‐printed scaffold‐perfusion bioreactor system are assessed on the production and bioactivity of EVs secreted from bone marrow‐derived mesenchymal stem cells (MSCs), a cell type widely implicated in generating EVs with therapeutic potential. The results indicate that perfusion bioreactor culture induces an ≈40‐80‐fold increase (depending on measurement method) in MSC EV production compared to conventional cell culture. Additionally, MSC EVs generated using the perfusion bioreactor system significantly improve wound healing in a diabetic mouse model, with increased CD31+ staining in wound bed tissue compared to animals treated with flask cell culture‐generated MSC EVs. Overall, this study establishes a promising solution to a major EV translational bottleneck, with the capacity for tunability for specific applications and general improvement alongside advancements in 3D‐printing technologies.

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Section: Introductionmentioning

confidence: 99%

Mesenchymal Stem Cell Culture within Perfusion Bioreactors Incorporating 3D‐Printed Scaffolds Enables Improved Extracellular Vesicle Yield with Preserved Bioactivity

Kronstadt

Patel

Born

et al. 2023

Adv Healthcare Materials

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“…Finally, storage and transport conditions of MSC-exosome products must also be addressed. Recently, several studies on the storage conditions and biological activity of the native and cargo-loading MSC-exosomes have carried out the ideal storage conditions (Levy et al, 2022). These results may suggest that lyophilization and storage at room temperature can preserve the enhanced bioactivity of cargo-loading exosomes.…”

Section: Prospects and Conclusionmentioning

confidence: 99%

“…In summary, various therapeutic molecules and nanoparticles can be efficiently transferred to exosomes using the abovementioned modification procedures. However, these methods require repeated purification and centrifugation using detergents, enzymes or other methods, which might disrupt the integrity of exosomes or reduce bioactivity (Levy et al, 2022). Therefore, novel bioengineering procedures are needed to compensate for these limitations.…”

Section: Direct Msc-exosome Engineeringmentioning

confidence: 99%

Bioengineered MSC-derived exosomes in skin wound repair and regeneration

Zhao

Wang

et al. 2023

Front. Cell Dev. Biol.

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Refractory skin defects such as pressure ulcers, diabetic ulcers, and vascular ulcers represent a challenge for clinicians and researchers in many aspects. The treatment strategies for wound healing have high cost and limited efficacy. To ease the financial and psychological burden on patients, a more effective therapeutic approach is needed to address the chronic wound. MSC-derived exosomes (MSC-exosomes), the main bioactive extracellular vesicles of the paracrine effect of MSCs, have been proposed as a new potential cell-free approach for wound healing and skin regeneration. The benefits of MSC-exosomes include their ability to promote angiogenesis and cell proliferation, increase collagen production, regulate inflammation, and finally improve tissue regenerative capacity. However, poor targeting and easy removability of MSC-exosomes from the wound are major obstacles to their use in clinical therapy. Thus, the concept of bioengineering technology has been introduced to modify exosomes, enabling higher concentrations and construction of particles of greater stability with specific therapeutic capability. The use of biomaterials to load MSC-exosomes may be a promising strategy to concentrate dose, create the desired therapeutic efficacy, and maintain a sustained release effect. The beneficial role of MSC-exosomes in wound healing is been widely accepted; however, the potential of bioengineering-modified MSC-exosomes remains unclear. In this review, we attempt to summarize the therapeutic applications of modified MSC-exosomes in wound healing and skin regeneration. The challenges and prospects of bioengineered MSC-exosomes are also discussed.

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“…All BEV samples were initially processed using centrifugation of culture followed by TFF of sterile culture supernatant, as detailed below. Additionally, after the terminal isolation procedure, all BEV samples were sterile filtered with a 0.2 μm syringe filter and stored at −20°C for no longer than 4 weeks until use, based on prior EV stability studies (Levy et al, 2023).…”

Section: Bacterial Culture and Productionmentioning

confidence: 99%

High performance anion exchange chromatography purification of probiotic bacterial extracellular vesicles enhances purity and anti‐inflammatory efficacy

Pirolli

Reus

Mamczarz

et al. 2023

Biotech & Bioengineering

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Bacterial extracellular vesicles (BEVs), including outer membrane vesicles, have emerged as a promising new class of vaccines and therapeutics to treat cancer and inflammatory diseases, among other applications. However, clinical translation of BEVs is hindered by a current lack of scalable and efficient purification methods. Here, we address downstream BEV biomanufacturing limitations by developing a method for orthogonal size‐ and charge‐based BEV enrichment using tangential flow filtration (TFF) in tandem with high performance anion exchange chromatography (HPAEC). The data show that size‐based separation coisolated protein contaminants, whereas size‐based TFF with charged‐based HPAEC dramatically improved purity of BEVs produced by probiotic Gram‐negative Escherichia coli and Gram‐positive lactic acid bacteria (LAB). Escherichia coli BEV purity was quantified using established biochemical markers while improved LAB BEV purity was assessed via observed potentiation of anti‐inflammatory bioactivity. Overall, this work establishes orthogonal TFF + HPAEC as a scalable and efficient method for BEV purification that holds promise for future large‐scale biomanufacturing of therapeutic BEV products.

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Impact of storage conditions and duration on function of native and cargo-loaded mesenchymal stromal cell extracellular vesicles

Cited by 15 publications

References 36 publications

Mesenchymal Stem Cell Culture within Perfusion Bioreactors Incorporating 3D‐Printed Scaffolds Enables Improved Extracellular Vesicle Yield with Preserved Bioactivity

Mesenchymal Stem Cell Culture within Perfusion Bioreactors Incorporating 3D‐Printed Scaffolds Enables Improved Extracellular Vesicle Yield with Preserved Bioactivity

Bioengineered MSC-derived exosomes in skin wound repair and regeneration

High performance anion exchange chromatography purification of probiotic bacterial extracellular vesicles enhances purity and anti‐inflammatory efficacy

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