Introducing micrometer-sized artificial objects into live cells: a method for cell-giant unilamellar vesicle...

Area of Science:

• Cell biology
• Biotechnology
• Nanotechnology

Background:

• Introducing large objects into mammalian cells is challenging.
• Existing methods often impact cell viability or efficiency.
• Giant unilamellar vesicles (GUVs) offer potential as carriers.

Purpose of the Study:

• To develop a method for introducing large objects (up to 1 µm) into mammalian cells.
• To assess the viability and proliferation of cells post-introduction.
• To demonstrate the potential for artificial symbiosis and applications in drug delivery and tissue engineering.

Main Methods:

• Preparation of GUVs containing artificial objects using a water-in-oil emulsion centrifugation method.
• Exposure of GUVs and HeLa cells to alternating current (AC) and direct current (DC) fields for electrofusion.
• Introduction of fluorescent beads (1 µm) and DNA origami nanostructures into cells.

Main Results:

• Successful and efficient introduction of 1 µm fluorescent beads and DNA origami into live mammalian cells.
• Electrofusion did not affect cell viability; cells proliferated normally.
• Introduced fluorescent beads were inherited during cell division, confirmed by confocal microscopy and flow cytometry.

Conclusions:

• The electrofusion of GUVs is a viable method for introducing large objects into mammalian cells.
• This technique supports normal cell proliferation and inheritance of introduced materials.
• The method holds significant potential for cell engineering, drug delivery, and tissue engineering applications.