Biofabrication of Cell-Derived Nanovesicles: A Potential Alternative to Extracellular Vesicles for Regenerative Medicine

Abstract

Extracellular vesicles (EVs) are mediators of intercellular communication by transferring functional biomolecules from their originating cells to recipient cells. This intrinsic ability has gained EVs increased scientific interest in their use as a direct therapeutic in the field of regenerative medicine or as vehicles for drug delivery. EVs derived from stem cells or progenitor cells can act as paracrine mediators to promote repair and regeneration of damaged tissues. Despite substantial research efforts into EVs for various applications, their use remains limited by the lack of highly efficient and scalable production methods. Here, we present the biofabrication of cell-derived nanovesicles (NVs) as a scalable, efficient, and cost-effective production alternative to EVs. We demonstrate that NVs have a comparable size and morphology as EVs, but lack standard EV (surface) markers. Additionally, in vitro uptake experiments show that human fetal cardiac fibroblast, endothelial cells, and cardiomyocyte progenitor cells internalize NVs. We observed that cardiac progenitor cell-derived NVs and EVs are capable of activating mitogen-activated protein kinase 1/2 (MAPK1/2)-extracellular signal-regulated kinase, and that both NVs and EVs derived from A431 and HEK293 cells can functionally deliver Cre-recombinase mRNA or protein to other cells. These observations indicate that NVs may have similar functional properties as EVs. Therefore, NVs have the potential to be applied for therapeutic delivery and regenerative medicine purposes.

Hijacking Endocytosis and Autophagy in Extracellular Vesicle Communication: Where the Inside Meets the Outside

“…Several studies have exploited the extremely sensitive CRE-based recombination technique to infer on EV-mediated trans-cytosol transport of biologically active macromolecules. Strikingly, these studies reveal that cytosolic entry of an EV cargo occurs at a sporadic rate (Ridder et al, 2014(Ridder et al, , 2015Zomer et al, 2015;Sterzenbach et al, 2017;Steenbeek et al, 2018;Ilahibaks et al, 2019), an observation in contradiction with the large body of evidence showing an efficient EV uptake through endocytosis.…”

Myocardial infarction affects Cx43 content of extracellular vesicles secreted by cardiomyocytes

“…The discrepancies among our in vivo, ex vivo, and in vitro data can be explained by various factors. For example, although our results demonstrate that ECM extracts were positive for CD63 and CD81 and devoid of intracellular contaminants, we cannot completely ensure that these vesicles do not partially result from artifactual release of intracellular contents during tissue processing, particularly during decellularization ( Théry et al, 2018 ; Ilahibaks et al, 2019 ).…”

Cell‐Derived Vesicles as TRPC1 Channel Delivery Systems for the Recovery of Cellular Respiratory and Proliferative Capacities

“…Sufficiently strong shear forces are capable of producing membrane vesicles, yet of relatively poorly defined composition due to the nonspecific nature of the shedding forces. [15][16][17] Inducing membrane blebbing with the use of cytochalasin B treatment alleviated the need for harsh mechanical disruption as well as improved CDV yield and functionality by allowing plasmamembrane signaling microdomains to remain intact. [8,9,18] Nonetheless, although CDVs produced in this manner are generally assumed to retain native plasma-membrane structure and integrity, detailed information of the proteome and liposome composition of CDVs remains sparse.…”