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
StatePublished - 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

Cite this

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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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.",
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Yoon, S, Kang, K, Cho, YD, Kim, Y, Buisson, EM, Yim, JH, Lee, SB, Ryu, K-Y, 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

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

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AU - Kang, Kyojin

AU - Cho, Young Duck

AU - Kim, Yohan

AU - Buisson, Elina Maria

AU - Yim, Ji Hye

AU - Lee, Seung Bum

AU - Ryu, Ki-Young

AU - Jeong, Jaemin

AU - Choi, Dongho

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