Technological advances towards extracellular vesicles mass production

Grangier, Alice; Branchu, Julien; Volatron, Jeanne; Piffoux, Max; Gazeau, Florence; Wilhelm, Claire; Silva, Amanda

doi:10.1016/j.addr.2021.113843

Cited by 97 publications

(77 citation statements)

References 121 publications

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“…The 3D culture conditions can optimize cell culture on a platform of limited size or scalability. It increases surface area while minimizing the volume of culture media per cell, consequently enhancing the concentration and yield of exosomes in conditioned media [ 15 ]. Gelma is one of the commonly used hydrogels simulating the physical structure and chemical composition of the natural extracellular matrix and providing a 3D template and extracellular matrix microenvironment [ 53 ].…”

Section: Discussionmentioning

confidence: 99%

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Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regeneration

et al. 2022

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Background Mesenchymal stem cells (MSCs) exosomes were previously shown to be effective in articular cartilage repairing. However, whether MSCs exosomes promote mature cartilage formation of microtia chondrocytes and the underlying mechanism of action remains unknown. Additionally, some hurdles, such as the low yield and unsatisfactory therapeutic effects of natural exosomes have emerged when considering the translation of exosomes-therapeutics to clinical practices or industrial production. Herein, we investigated the roles of human adipose-derived stem cells (ADSCs) exosomes in modulating microtia chondrocytes and the underlying mechanism of action. Special attention was also paid to the mass production and functional modification of ADSCs exosomes. Results We firstly used porous gelatin methacryloyl (Porous Gelma) hydrogel with pores size of 100 to 200 μm for 3D culture of passage 2, 4 and 6 ADSCs (P2, P4 and P6 ADSCs, respectively), and obtained their corresponding exosomes (Exo 2, Exo 4 and Exo 6, respectively). In vitro results showed Exo 2 outperformed both Exo 4 and Exo 6 in enhancing cell proliferation and attenuating apoptosis. However, both Exo 4 and Exo 6 promoted chondrogenesis more than Exo 2 did. Small RNA sequencing results indicated Exo 4 was similar to Exo 6 in small RNA profiles and consistently upregulated PI3K/AKT/mTOR signaling pathway. Notably, we found hsa-miR-23a-3p was highly expressed in Exo 4 and Exo 6 compared to Exo 2, and they modulated microtia chondrocytes by transferring hsa-miR-23a-3p to suppress PTEN expression, and consequently to activate PI3K/AKT/mTOR signaling pathway. Then, we designed genetically engineered exosomes by directly transfecting agomir-23a-3p into parent P4 ADSCs and isolated hsa-miR-23a-3p-rich exosomes for optimizing favorable effects on cell viability and new cartilage formation. Subsequently, we applied the engineered exosomes to in vitro and in vivo tissue-engineered cartilage culture and consistently found that the engineered exosomes could enhance cell proliferation, attenuate apoptosis and promote cartilage regeneration. Conclusions Taken together, the porous Gelma hydrogel could be applied to exosomes mass production, and functional modification could be achieved by selecting P4 ADSCs as parent cells and genetically modifying ADSCs. Our engineered exosomes are a promising candidate for tissue-engineered ear cartilage regeneration. Graphical Abstract

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Section: Discussionmentioning

confidence: 99%

“…Indeed, hurdles have concurrently emerged when considering the translation of exosomes-therapeutics to clinical practices or industrial production [ 15 ]. The first one is the low yield, which is mainly attributed to the poor secretion from parent MSCs, the limited culture space and inefficient isolation methods.…”

Section: Introductionmentioning

confidence: 99%

Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regeneration

et al. 2022

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“…For EVs mass production, there are two main development directions. One is to optimize the culture of source cells, and the other is to find more suitable stimulation to increase EV production per cell [ 120 ]. Multiple bioreactors like “flask” bioreactor and stirred tank bioreactors were designed for large-scale culture of source cells [ 121 – 123 ].…”

Section: Perspectives and Challengesmentioning

confidence: 99%

Engineered extracellular vesicles: potentials in cancer combination therapy

et al. 2022

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Extracellular vesicles (EVs) are a group of secretory vesicles with cell-derived membrane and contents. Due to the cargo delivery capability, EVs can be designed as drug delivery platforms for cancer therapy. Biocompatibility and immune compatibility endow EVs with unique advantages compared with other nanocarriers. With the development of this field, multiple ingenious modification methods have been developed to obtain engineered EVs with desired performance. Application of engineered EVs in cancer therapy has gradually shifted from monotherapy to combinational therapy to fight against heterogeneous cancer cells and complex tumor microenvironment. In addition, the strong plasticity and load capacity of engineered EV make it potential to achieve various combinations of cancer treatment methods. In this review, we summarize the existing schemes of cancer combination therapy realized by engineered EVs, highlight the mechanisms and representative examples of these schemes and provide guidance for the future application of engineered EVs to design more effective cancer combination treatment plans. Graphical Abstract

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“…In the present work, we focus on the isolation of nanoalgosomes and we optimize an efficient bioprocess for a sustainable, scalable, and renewable EVs production, along with a robust quality control procedure, as defined in our previous work ( Adamo et al, 2021 ) in accord to the guidelines and the consensus from the scientific community ( Théry et al, 2018 ). A cost-effective and reliable EVs production, which is also suitable for an industrial or large scale exploitation, requires a fine tuning of both upstream and downstream processes ( Paganini et al, 2019 ; Buschmann et al, 2021 ; Grangier et al, 2021 ; Staubach et al, 2021 ). Here, we discuss and define a clear manufacturing practice for the implementation of nanoalgosome production, with optimized protocols for microalgal cultivation ( upstream processing ) and isolation of EVs by Tangential Flow Filtration (TFF), an isolation technique allowing to process large volumes of microalgae cultures, reaching concentrated EV samples ( downstream processing ).…”

Section: Introductionmentioning

confidence: 99%

Isolation of Extracellular Vesicles From Microalgae: A Renewable and Scalable Bioprocess

Paterna

Rao

Adamo

et al. 2022

Front. Bioeng. Biotechnol.

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Extracellular vesicles (EVs) play a crucial role as potent signal transducers among cells, with the potential to operate cross-species and cross-kingdom communication. Nanoalgosomes are a subtype of EVs recently identified and isolated from microalgae. Microalgae represent a natural bioresource with the capacity to produce several secondary metabolites with a broad range of biological activities and commercial applications. The present study highlights the upstream and downstream processes required for the scalable production of nanoalgosomes from cultures of the marine microalgae Tetraselmis chuii. Different technical parameters, protocols, and conditions were assessed to improve EVs isolation by tangential flow filtration (TFF), aiming to enhance sample purity and yield. The optimization of the overall bioprocess was enhanced by quality control checks operated through robust biophysical and biochemical characterizations. Further, we showed the possibility of recycling by TFF microalgae cells post-EVs isolation for multiple EV production cycles. The present results highlight the potential of nanoalgosome production as a scalable, cost-effective bioprocess suitable for diverse scientific and industrial exploitations.

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Technological advances towards extracellular vesicles mass production

Cited by 97 publications

References 121 publications

Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regeneration

Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regeneration

Engineered extracellular vesicles: potentials in cancer combination therapy

Isolation of Extracellular Vesicles From Microalgae: A Renewable and Scalable Bioprocess

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