Introducing micrometer-sized artificial objects into live cells

A method for cell-giant unilamellar vesicle electrofusion

Akira C. Saito, Toshihiko Ogura, Kei Fujiwara, Satoshi Murata, Shin Ichiro M Nomura

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Here, we report a method for introducing large objects of up to a micrometer in diameter into cultured mammalian cells by electrofusion of giant unilamellar vesicles. We prepared GUVs containing various artificial objects using a water-in-oil (w/o) emulsion centrifugation method. GUVs and dispersed HeLa cells were exposed to an alternating current (AC) field to induce a linear cell-GUV alignment, and then a direct current (DC) pulse was applied to facilitate transient electrofusion. With uniformly sized fluorescent beads as size indexes, we successfully and efficiently introduced beads of 1 μm in diameter into living cells along with a plasmid mammalian expression vector. Our electrofusion did not affect cell viability. After the electrofusion, cells proliferated normally until confluence was reached, and the introduced fluorescent beads were inherited during cell division. Analysis by both confocal microscopy and flow cytometry supported these findings. As an alternative approach, we also introduced a designed nanostructure (DNA origami) into live cells. The results we report here represent a milestone for designing artificial symbiosis of functionally active objects (such as micro-machines) in living cells. Moreover, our technique can be used for drug delivery, tissue engineering, and cell manipulation.

Original languageEnglish
Article numbere106853
JournalPLoS One
Volume9
Issue number9
DOIs
Publication statusPublished - 2014 Sep 17

Fingerprint

electrofusion
Unilamellar Liposomes
giant cells
Giant Cells
Cells
cells
methodology
Flow cytometry
Centrifugation
Confocal microscopy
Emulsions
Drug delivery
Symbiosis
Tissue engineering
Nanostructures
Tissue Engineering
HeLa Cells
Oils
Confocal Microscopy
Plasmids

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Introducing micrometer-sized artificial objects into live cells : A method for cell-giant unilamellar vesicle electrofusion. / Saito, Akira C.; Ogura, Toshihiko; Fujiwara, Kei; Murata, Satoshi; Nomura, Shin Ichiro M.

In: PLoS One, Vol. 9, No. 9, e106853, 17.09.2014.

Research output: Contribution to journalArticle

Saito, Akira C. ; Ogura, Toshihiko ; Fujiwara, Kei ; Murata, Satoshi ; Nomura, Shin Ichiro M. / Introducing micrometer-sized artificial objects into live cells : A method for cell-giant unilamellar vesicle electrofusion. In: PLoS One. 2014 ; Vol. 9, No. 9.
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