Nonintegrating direct conversion using mRNA into hepatocyte-like cells

Sangtae Yoon, Kyojin Kang, Young Duck Cho, Yohan Kim, Elina Maria Buisson, Ji Hye Yim, Seung Bum Lee, Ki Young Ryu, Jaemin Jeong, Dongho Choi

Research output: Contribution to journalArticle

Abstract

Recently, several researchers have reported that direct reprogramming techniques can be used to differentiate fibroblasts into hepatocyte-like cells without a pluripotent intermediate step. However, the use of viral vectors for conversion continues to pose important challenges in terms of genome integration. Herein, we propose a new method of direct conversion without genome integration with potential clinical applications. To generate hepatocyte-like cells, mRNA coding for the hepatic transcription factors Foxa3 and HNF4α was transfected into mouse embryonic fibroblasts. After 10-12 days, the fibroblasts converted to an epithelial morphology and generated colonies of hepatocyte-like cells (R-iHeps). The generated R-iHeps expressed hepatocyte-specific marker genes and proteins, including albumin, alpha-fetoprotein, HNF4α, CK18, and CYP1A2. To evaluate hepatic function, indocyanine green uptake, periodic acid-Schiff staining, and albumin secretion were assessed. Furthermore, mCherry-positive R-iHeps were engrafted in the liver of Alb-TRECK/SCID mice, and we confirmed FAH enzyme expression in Fah1RTyrc/RJ models. In conclusion, our data suggest that the nonintegrating method using mRNA has potential for cell therapy.

Original languageEnglish
Article number8240567
JournalBioMed Research International
Volume2018
DOIs
Publication statusPublished - 2018 Jan 1

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Hepatocytes
Fibroblasts
Messenger RNA
Albumins
Liver
Hepatocyte Nuclear Factor 4
Genes
Genome
Cytochrome P-450 CYP1A2
Periodic Acid
Indocyanine Green
SCID Mice
alpha-Fetoproteins
Cell- and Tissue-Based Therapy
Transcription Factors
Research Personnel
Staining and Labeling
Enzymes
Proteins

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Yoon, S., Kang, K., Cho, Y. D., Kim, Y., Buisson, E. M., Yim, J. H., ... Choi, D. (2018). Nonintegrating direct conversion using mRNA into hepatocyte-like cells. BioMed Research International, 2018, [8240567]. https://doi.org/10.1155/2018/8240567

Nonintegrating direct conversion using mRNA into hepatocyte-like cells. / Yoon, Sangtae; Kang, Kyojin; Cho, Young Duck; Kim, Yohan; Buisson, Elina Maria; Yim, Ji Hye; Lee, Seung Bum; Ryu, Ki Young; Jeong, Jaemin; Choi, Dongho.

In: BioMed Research International, Vol. 2018, 8240567, 01.01.2018.

Research output: Contribution to journalArticle

Yoon, S, Kang, K, Cho, YD, Kim, Y, Buisson, EM, Yim, JH, Lee, SB, Ryu, KY, Jeong, J & Choi, D 2018, 'Nonintegrating direct conversion using mRNA into hepatocyte-like cells', BioMed Research International, vol. 2018, 8240567. https://doi.org/10.1155/2018/8240567
Yoon, Sangtae ; Kang, Kyojin ; Cho, Young Duck ; Kim, Yohan ; Buisson, Elina Maria ; Yim, Ji Hye ; Lee, Seung Bum ; Ryu, Ki Young ; Jeong, Jaemin ; Choi, Dongho. / Nonintegrating direct conversion using mRNA into hepatocyte-like cells. In: BioMed Research International. 2018 ; Vol. 2018.
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