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Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cells.


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 HNF4? 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, HNF4?, 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.

SUBMITTER: Yoon S 

PROVIDER: S-EPMC6171260 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cells.

Yoon Sangtae S   Kang Kyojin K   Cho Young-Duck YD   Kim Yohan Y   Buisson Elina Maria EM   Yim Ji-Hye JH   Lee Seung Bum SB   Ryu Ki-Young KY   Jeong Jaemin J   Choi Dongho D  

BioMed research international 20180920


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 transcrip  ...[more]

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