Efficient mRNA delivery with graphene oxide-polyethylenimine for generation of footprint-free human induced pluripotent stem cells

Clinical applications of induced pluripotent stem cells (iPSCs) require development of technologies for the production of “footprint-free” (gene integration-free) iPSCs, which avoid the potential risk of insertional mutagenesis in humans. Previously, several studies have shown that mRNA transfer can...

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Bibliographic Details
Published in:Journal of controlled release 2016-08, Vol.235, p.222-235
Main Authors: Choi, Hye Yeon, Lee, Tae-Jin, Yang, Gwang-Mo, Oh, Jaesur, Won, Jihye, Han, Jihae, Jeong, Gun-Jae, Kim, Jongpil, Kim, Jin-Hoi, Kim, Byung-Soo, Cho, Ssang-Goo
Format: Article
Language:English
Subjects:
Adipose Tissue - cytology
Alkaline Phosphatase - metabolism
Cell Differentiation - drug effects
Cell Survival - drug effects
Fibroblasts - cytology
Footprint-free transgene-free
Gene delivery
Graphene oxide-polyethylenimine complex
Graphite - administration & dosage
HEK293 Cells
Human induced pluripotent stem cells
Humans
Induced Pluripotent Stem Cells - cytology
Integration-free
iPSC
Oxides - administration & dosage
Polyethyleneimine - administration & dosage
RNA delivery
RNA, Messenger - administration & dosage
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Summary:Clinical applications of induced pluripotent stem cells (iPSCs) require development of technologies for the production of “footprint-free” (gene integration-free) iPSCs, which avoid the potential risk of insertional mutagenesis in humans. Previously, several studies have shown that mRNA transfer can generate “footprint-free” iPSCs, but these studies did not use a delivery vehicle and thus repetitive daily transfection was required because of mRNA degradation. Here, we report an mRNA delivery system employing graphene oxide (GO)-polyethylenimine (PEI) complexes for the efficient generation of “footprint-free” iPSCs. GO-PEI complexes were found to be very effective for loading mRNA of reprogramming transcription factors and protection from mRNA degradation by RNase. Dynamic suspension cultures of GO-PEI/RNA complexes-treated cells dramatically increased the reprogramming efficiency and successfully generated rat and human iPSCs from adult adipose tissue-derived fibroblasts without repetitive daily transfection. The iPSCs showed all the hallmarks of pluripotent stem cells including expression of pluripotency genes, epigenetic reprogramming, and differentiation into the three germ layers. These results demonstrate that mRNA delivery using GO-PEI-RNA complexes can efficiently generate “footprint-free” iPSCs, which may advance the translation of iPSC technology into the clinical settings. [Display omitted]
ISSN:0168-3659
1873-4995
DOI:10.1016/j.jconrel.2016.06.007