Mesenchymal stem cell culture within perfusion bioreactors incorporating 3D-printed scaffolds enables improved extracellular vesicle yield with preserved bioactivity

Kronstadt, Stephanie M.; Patel, Divya B.; Born, Louis J.; Levy, Daniel; Lerman, Max J.; Mahadik, Bhushan; McLoughlin, Shannon Theresa; Fasuyi, Arafat; Fowlkes, Lauren; Heyningen, Lauren Hoorens Van; Aranda, Amaya; Abadchi, Sanaz Nourmohammadi; Chang, Kyungho; Hsu, An; Bengali, Sameer; Harmon, John W.; Fisher, John P.; Jay, Steven M.

doi:10.1101/2022.08.30.505860

Cited by 3 publications

(2 citation statements)

References 68 publications

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“…Novel investigations have used more straightforward methods (such as flat-plate bioreactors), permitting a more accurate approach for comprehend and restraining flow-derived shear stress. However, still absence the adjustability essential for dynamic construction and experimental investigation requirements [69]. Moreover, stirred suspension bioreactors (SSBs) have been efficiently utilized to scale up MSCs manufacture.…”

Section: Msc-exos Produced By Bioreactor Production Methodsmentioning

confidence: 99%

The potential use of mesenchymal stem cells-derived exosomes as microRNAs delivery systems in different diseases

Oveili¹,

Vafaei

Bazavar

et al. 2023

Cell Commun Signal

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MicroRNAs (miRNAs) are a group of small non-coding RNAs that regulate gene expression by targeting mRNA. Moreover, it has been shown that miRNAs expression are changed in various diseases, such as cancers, autoimmune disease, infectious diseases, and neurodegenerative Diseases. The suppression of miRNA function can be easily attained by utilizing of anti-miRNAs. In contrast, an enhancement in miRNA function can be achieved through the utilization of modified miRNA mimetics. The discovery of appropriate miRNA carriers in the body has become an interesting subject for investigators. Exosomes (EXOs) therapeutic efficiency and safety for transferring different cellular biological components to the recipient cell have attracted significant attention for their capability as miRNA carriers. Mesenchymal stem cells (MSCs) are recognized to generate a wide range of EXOs (MSC-EXOs), showing that MSCs may be effective for EXO generation in a clinically appropriate measure as compared to other cell origins. MSC-EXOs have been widely investigated because of their immune attributes, tumor-homing attributes, and flexible characteristics. In this article, we summarized the features of miRNAs and MSC-EXOs, including production, purification, and miRNA loading methods of MSC-EXOs, and the modification of MSC-EXOs for targeted miRNA delivery in various diseases. Graphical abstract

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Section: Msc-exos Produced By Bioreactor Production Methodsmentioning

confidence: 99%

The potential use of mesenchymal stem cells-derived exosomes as microRNAs delivery systems in different diseases

Oveili¹,

Vafaei

Bazavar

et al. 2023

Cell Commun Signal

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show abstract

“…For instance, the impact of a 3D-printed scaffold-perfusion bioreactor system on the production and bioactivity of EVs secreted by MSCs has been explored. Findings suggest that culture in a perfusion bioreactor can yield an approximate 40-80-fold increase in MSC EVs production, varying with the measurement technique, compared to traditional cell culture methods [79]. Although the application of MSC EVs produced via this system in liver disease has not yet been fully evaluated, the approach holds substantial potential for future therapeutic interventions.…”

Section: Scaling-up Isolation and Bioengineering Of Clinical-grade Ms...mentioning

confidence: 99%

Mesenchymal stromal cell-derived extracellular vesicles therapy openings new translational challenges in immunomodulating acute liver inflammation

Sitbon,

Delmotte,

Pistorio

et al. 2024

J Transl Med

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Inflammation plays a critical role in conditions such as acute liver failure, acute-on-chronic liver failure, and ischemia–reperfusion-induced liver injury. Various pathogenic pathways contribute to liver inflammation, involving inflammatory polarization of macrophages and Küpffer cells, neutrophil infiltration, dysregulation of T cell subsets, oxidative stress, and activation of hepatic stellate cells. While mesenchymal stromal cells (MSCs) have demonstrated beneficial properties, their clinical translation is limited by their cellular nature. However, MSC-derived extracellular vesicles (MSC-EVs) have emerged as a promising cell-free therapeutic approach for immunomodulation. MSC-EVs naturally mirror their parental cell properties, overcoming the limitations associated with the use of MSCs. In vitro and in vivo preclinical studies have demonstrated that MSC-EVs replicate the beneficial effects of MSCs in liver injury. This includes the reduction of cell death and oxidative stress, improvement of hepatocyte function, induction of immunomodulatory effects, and mitigation of cytokine storm. Nevertheless, MSC-EVs face challenges regarding the necessity of defining consistent isolation methods, optimizing MSCs culture conditions, and establishing quality control measures for EV characterization and functional assessment. By establishing standardized protocols, guidelines, and affordable cost mass production, clinicians and researchers will have a solid foundation to conduct further studies, validate the therapeutic efficacy of MSC-EVs, and ultimately pave the way for their clinical implementation in acute liver injury.

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Dynamic cultivation of human mesenchymal stem/stromal cells for the production of extracellular vesicles in a 3D bioreactor system

Almeria,

Weiss,

Keck

et al. 2024

Biotechnol Lett

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Purpose 3D cell culture and hypoxia have been demonstrated to increase the therapeutic effects of mesenchymal stem/stromal cells (MSCs)-derived extracellular vesicles (EVs). In this study, a process for the production of MSC-EVs in a novel 3D bioreactor system under normoxic and hypoxic conditions was established and the resulting EVs were characterized. Methods Human adipose-derived MSCs were seeded and cultured on a 3D membrane in the VITVO® bioreactor system for 7 days. Afterwards, MSC-EVs were isolated and characterized via fluorescence nanoparticle tracking analysis, flow cytometry with staining against annexin V (Anx5) as a marker for EVs exposing phosphatidylserine, as well as CD73 and CD90 as MSC surface markers. Results Cultivation of MSC in the VITVO® bioreactor system demonstrated a higher concentration of MSC-EVs from the 3D bioreactor (9.1 × 109 ± 1.5 × 109 and 9.7 × 109 ± 3.1 × 109 particles/mL) compared to static 2D culture (4.2 × 109 ± 7.5 × 108 and 3.9 × 109 ± 3.0 × 108 particles/mL) under normoxic and hypoxic conditions, respectively. Also, the particle-to-protein ratio as a measure for the purity of EVs increased from 3.3 × 107 ± 1.1 × 107 particles/µg protein in 2D to 1.6 × 108 ± 8.3 × 106 particles/µg protein in 3D. Total MSC-EVs as well as CD73−CD90+ MSC-EVs were elevated in 2D normoxic conditions. The EV concentration and size did not differ significantly between normoxic and hypoxic conditions. Conclusion The production of MSC-EVs in a 3D bioreactor system under hypoxic conditions resulted in increased EV concentration and purity. This system could be especially useful in screening culture conditions for the production of 3D-derived MSC-EVs.

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Mesenchymal stem cell culture within perfusion bioreactors incorporating 3D-printed scaffolds enables improved extracellular vesicle yield with preserved bioactivity

Cited by 3 publications

References 68 publications

The potential use of mesenchymal stem cells-derived exosomes as microRNAs delivery systems in different diseases

The potential use of mesenchymal stem cells-derived exosomes as microRNAs delivery systems in different diseases

Mesenchymal stromal cell-derived extracellular vesicles therapy openings new translational challenges in immunomodulating acute liver inflammation

Dynamic cultivation of human mesenchymal stem/stromal cells for the production of extracellular vesicles in a 3D bioreactor system

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