Abstract
Despite huge need in the medical domain and significant development efforts, artificial cells to date have limited composition and functionality. Although some artificial cells have proven successful for producing therapeutics or performing in vitro specific reactions, they have not been investigated in vivo to determine whether they preserve their architecture and functionality while avoiding toxicity. Here, these limitations are overcome and customizable cell mimic is achieved—molecular factories (MFs)—by supplementing giant plasma membrane vesicles derived from donor cells with nanometer‐sized artificial organelles (AOs). MFs inherit the donor cell's natural cytoplasm and membrane, while the AOs house reactive components and provide cell‐like architecture and functionality. It is demonstrated that reactions inside AOs take place in a close‐to‐nature environment due to the unprecedented level of complexity in the composition of the MFs. It is further demonstrated that in a zebrafish vertebrate animal model, these cell mimics show no apparent toxicity and retain their integrity and function. The unique advantages of highly varied composition, multicompartmentalized architecture, and preserved functionality in vivo open new biological avenues ranging from the study of biorelevant processes in robust cell‐like environments to the production of specific bioactive compounds.
Highlights
Despite huge need in the medical domain and significant development microenvironments and compartmentalized spaces
These limitations are overcome functional molecular units with supramolecular assemblies to develop the first prototypes of artificial cells[1,2] with the aim of providing micrometer range compartments with and customizable cell mimic is achieved—molecular factories (MFs)—by cell-like properties for understanding supplementing giant plasma membrane vesicles derived from donor cells fundamental biological phenomena and with nanometer-sized artificial organelles (AOs)
We have introduced customizable MFs as cell mimics that have a natural membrane and cytosol interior combined with AOs that produce desired molecules or signals
Summary
Achievement of a close-to-nature cell mimic requires the transfer of functional elements (confined AOs, synthetic- and biomolecules) necessary to support overall functionality inside GPMVs used as micrometer sized compartments with intrinsic natural composition.[2]. The components of the vesiculation buffer are not expected to show toxicity in vivo after E-GPMV formation, due to their relatively low half-life.[32] In order to establish the size distribution of E-GPMVs, we supplement them with a fluorescent model membrane protein LcK tyrosine kinaseGFP (Lck-GFP) by genetically engineering HepG2 cells to overexpress this protein using a baculovirus gene transfer (BacMam 2.0).[33] When Lck-GFP is successfully inserted, it preserves its fluorescence property, and does not affect the integrity of the membrane compartment (Figure S1, Supporting Information), in agreement with previous reports about the insertion of gap junction proteins in the membrane of giant plasma membrane vesicles.[26,34,35] E-GPMVs are isolated with a size range of 2–20 μm, as demonstrated by confocal laser scanning microscopy (CLSM) and flow cytometry data (Figure 2A–C).
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