Review: MSCs and Exosomes Production

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www.pelobiotech.com breast milk. MSCs have further been defined by the International Society for Cellular Therapy (ISCT) based on specific criteria outlined in a position statement from 2006. According to these criteria, MSCs must exhibit certain characteristics to be classified as such. These include the ability to adhere to plastic surfaces during in vitro culture, expression of specific surface markers such as CD105, CD73, and CD90 while lacking CD45, CD34, CD14, or CD11b, expression markers and the capacity to differentiate into osteoblasts, adipocytes, and chondrocytes under specific in vitro conditions 22,23 . These criteria are crucial for the identification and standardization of MSCs across various tissue sources. Importantly, the tissue source of MSCs can influence their therapeutic potential, making it essential to understand the differences between MSCs isolated from different tissues to predict their behaviour and widen their clinical use 1 . Extracellular Vesicles: Nature's Nanoscale Messengers EVs, nanoscale membrane-bound vesicles released by various cell types, including MSCs, play a vital role in an array of cellular functions, including intercellular communication, cell differentiation, and proliferation, angiogenesis, stress response, and immune signalling. The ability to carry out these different functions is because of the complexity of EVs. These vesicles carry and transfer functional cargo like proteins, messenger RNAs, microRNAs, cytokines, lipids, cell surface receptors, enzymes, and transcription factors from cells to the recipient cells. Their sizes range from 30 to 150 nanometres, originating from a specialized biogenesis pathway. The composition of EVs is contingent on the donor cell type and the physiological context of production. EVs interact with recipient cells through specific adhesion molecules and can be internalized via multiple pathways, dependent on the proximity of the target cells. The nucleic acid content in EVs is particularly influential in their functional capacity. They harbour distinct markers including tetraspanins (CD9, CD63, CD81), integrins, MHC molecules, HSP70, HSP90, Alix, TSG101, and GTPases. The lipid bilayer encapsulating EVs confers stability and protection, facilitating their biological roles. MSCs release large amounts of EVs for cell-tocell communication, maintaining a dynamic and homeostatic microenvironment for tissue repair and regeneration 2,28 . Furthermore, EVs have been implicated in various physiological and pathological processes, including cardiovascular diseases and neurogenesis 3,29-31 . Tel: +49 (0) 89 517 286 59 0 Mail: info@pelobiotech.com
www.pelobiotech.com MSC-Derived EVs: Key Agents in Regenerative Medicine Past research has demonstrated that, despite positive effects in various settings, MSCs were barely detected in affected tissues, resulting in the hypothesis that they mainly act via their secretome rather than in a direct cellular manner 15 . Using the examples of an acute kidney injury model and a myocardial infarction model that MSCs were found to exert their therapeutic effects EVs 33 . MSC-derived EVs have become key players in regenerative medicine, showcasing a broad range of therapeutic effects. Originating from MSCs, these EVs carry immunomodulatory, regenerative, and anti-inflammatory traits, making them highly effective in tissue repair, angiogenesis, inflammation control, and wound healing. They contribute significantly to critical cellular processes, including angiogenesis, fibrosis reduction, and remodelling of the extracellular matrix. They are particularly promising for treating a spectrum of conditions such as cardiovascular, renal, hepatic, pulmonary, and neurodegenerative diseases, and they also exhibit antimicrobial effects. Unlike MSCs themselves, which can pose challenges related to cell viability, potential for immune rejection, and the complexity of direct use in regenerative therapies, MSC-EVs offer a safer, more stable, and potentially more effective alternative. In contrast to cell therapies, EVs are not self-replicating, and they lack endogenous tumour formation potentials. EVs do not seem to sense environmental conditions, and thus, their biological activity can be predicted more reliably than that of cells. Preconditioning and engineering techniques have enhanced the efficacy of MSC-EVs, paving the way for improved outcomes in cell-free therapeutic interventions 8 . This evolution emphasizes the strategic advantage of utilizing MSC-EVs over direct MSC therapies, as they represent an innovative method for harnessing the full regenerative potential of mesenchymal stem cells, setting a new standard in medical treatments. Culture and Expansion of MSCs for Extracellular Vesicle Production The origin, culture, and expansion of MSCs are crucial for EVs production. The choice of expansion method significantly impacts EVs yield and quality. The field of MSC research faces challenges due to the inherent tendency of primary MSCs to undergo senescence during culture expansion. This limitation has prompted researchers to explore the generation and characterization of immortalized MSC (iMSC) lines as a potential solution. IMSC lines, such as those created by inducing the expression of human telomerase reverse transcriptase (hTERT), have been investigated for their ability to offer a reliable and scalable source of MSCs for EVs production 17 . Studies have indicated that iMSC lines could serve as a consistent resource for EVs production, which is crucial for various therapeutic applications. However, it is important to note that iMSCs may exhibit functional alterations compared to primary MSCs 18 . Tel: +49 (0) 89 517 286 59 0 Mail: info@pelobiotech.com
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Review: MSCs and Exosomes Production

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