Abstract
Nanoscale biocomponents naturally released by cells, such as extracellular vesicles (EVs), have recently gained interest due to their therapeutic and diagnostic potential. Membrane based isolation and co-culture systems have been utilized in an effort to study EVs and their effects. Nevertheless, improved platforms for the study of small EVs are still needed. Suitable membranes, for isolation and co-culture systems, require pore sizes to reach into the nanoscale. These pore sizes cannot be achieved through traditional lithographic techniques and conventional thick nanoporous membranes commonly exhibit low permeability. Here we utilized nanospheres, similar in size and shape to the targeted small EVs, as patterning features for the fabrication of freestanding SiN membranes (120 nm thick) released in minutes through a sacrificial ZnO layer. We evaluated the feasibility of separating a subpopulation of EVs based on size using these membranes. The membrane used here showed an effective size cut-off of 300 nm with the majority of the EVs ≤200 nm. This work provides a convenient platform with great potential for studying subpopulations of EVs.
Highlights
Cells are known to secrete extracellular vesicles (EVs) ranging in size from 30 to 1000 nm.[1]
All substrates used were silicon wafers with h100i orientation purchased from University Wafer (Massachusetts, United States) coated with 100 nm of zinc oxide (ZnO) deposited at room temperature from a Zn target purchased from Kurt J
As the substrate travels through an interface, several parameters come into play to adequately balance attractive and repulsive forces.[56,71]
Summary
Cells are known to secrete extracellular vesicles (EVs) ranging in size from 30 to 1000 nm.[1]. We have demonstrated the viability of utilizing those membranes for cell culture studies.[38,54] In the current work, we utilize a patterning approach with close control of pore size over the range of small EVs, together with a simple methodology for the release and integration of the ultrathin membrane into a silicone based device for the study of EVs. To the best of our knowledge, such a comprehensive process has not yet been demonstrated.
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