Activation of pluripotency genes by a nanotube‐mediated protein delivery system

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Summary: The overexpression of cell reprogramming factors (Oct4, Sox2, Klf4, Nanog, and c‐Myc) allows differentiated cells to revertto an earlier developmental stage. Differentiated cells can also be reprogrammed by directly delivering reprogramming proteins tagged with cell‐penetrating peptides, which allow the proteins to pass through the cell membrane and into the cytoplasm–although this method has been an inefficient process. Here, we describe a novel technique for delivering reprogramming proteins into cells using titanium oxide (TiO2) nanotubes, which show no cytotoxic effects and do not affect cell proliferation. TiO2 nanotubes successfully transferred the above‐mentioned reprogramming factors into differentiated somatic cells. After 3 weeks of treatment with protein‐conjugated nanotubes, the somatic cells adopted an embryonic stem cell‐like morphology and expressed activated Oct4‐green fluorescent protein, a pluripotency biomarker. Our results indicate that TiO2 nanotubes can be used to directly deliver reprogramming factors into somatic cells to induce pluripotency. Mol. Reprod. Dev. 80: 1000–1008, 2013. © 2013 Wiley Periodicals, Inc.

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