Designer Extracellular Vesicles Modulate Pro‐Neuronal Cell Responses and Improve Intracranial Retention (Adv. Healthcare Mater. 5/2022)

Abstract

Functionalized Extracellular Vesicles In article number 2100805 by Natalia Higuita‐Castro and co‐workers, designer extracellular vesicles (EVs) are manufactured via non‐viral engineering of donor cells through electroporation. Designer EVs are packed with plasmid DNA, mRNA, and protein content. Transfecting donor cells with expression plasmids for membrane proteins can be used to tailor the surface decoration of the EVs. Pro‐neurogenic designer EVs drive neurogenesis and improve targeting of neuronal tissue, in vitro and in vivo.

Engineered Extracellular Vesicles from Human Skin Cells Induce Pro‐β‐Cell Conversions in Pancreatic Ductal Cells

“…We have previously shown that the engineered cargo of the EVs is primarily made of copies of the transfected expression plasmids as well as mRNA transcripts and proteins of the same factors. [ 14,16,21 ] qRT‐PCR analysis of PNM‐loaded EVs pretreated with DNAse confirms the absence of plasmid DNA coprecipitates in the EV preparations (Figure S2A, Supporting Information). Overall, peak PNM expression in the donor cells and loading in the EVs were found as early as 6 h post‐transfection.…”

“…CryoEM grids were prepared as previously described [ 14 ] by applying a small aliquot (3 μL) of sample to the carbon side of Lacey carbon grid. After blotting away excess liquid, the grid was immediately plunged into liquid ethane to rapidly form a thin film of amorphous ice using Vitrobot Mark IV system (Thermo Fisher Scientific, Hillsboro).…”

“…Extracellular vesicles (EVs) have recently emerged as promising nonviral gene carriers for different applications. [ 14–24 ] EVs are biological nanoparticles produced by all cell types, which contain a wide variety of molecular cargo (e.g., mRNA, micro‐RNA, and protein), and are involved in mediating short‐ and long‐range intercellular communication. [ 25 ] EVs have been shown to offer many advantages over more standard nanocarrier systems, including high stability in circulation and the capacity to carry larger molecular cargo.…”

“…Thus, the implementation of EV‐based strategies to deploy β cell‐directed reprogramming factors, in vivo, could potentially overcome many of the aforementioned limitations. We have previously shown that EVs could serve as effective carrier systems to drive different types of reprogramming processes in vitro and in vivo; [ 14,15,21,23 ] however, the ability of EVs to deploy and drive β cell‐directed reprogramming signals, in vivo, remains unknown.…”