Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cells

• Published in: BioMed research international 2018-01, Vol.2018 (2018), p.1-8
• Main Authors: Choi, Dongho, Jeong, Jaemin, Lee, Seung Bum, Yim, Ji-Hye, Buisson, Elina Maria, Kim, Yohan, Cho, Young-duck, Kang, Kyojin, Yoon, Sangtae, Ryu, Ki-Young
• Format: Article
• Language: English
• Subjects: Albumin; Animal models; Animals; Antigens, Differentiation - biosynthesis; Antigens, Differentiation - genetics; Cell Differentiation; CYP1A2 protein; Cytochrome P450; Direct conversion; DNA binding proteins; Embryo fibroblasts; Embryo, Mammalian - cytology; Embryo, Mammalian - metabolism; Embryos; Enzymes; Fibroblasts; Fibroblasts - cytology; Fibroblasts - metabolism; Genomes; Growth factors; Hepatocyte Nuclear Factor 3-gamma - biosynthesis; Hepatocyte Nuclear Factor 3-gamma - genetics; Hepatocyte Nuclear Factor 4 - genetics; Hepatocytes - cytology; Hepatocytes - metabolism; Integration; Liver; Liver cancer; Liver diseases; Messenger RNA; Metabolism; Mice; Mice, SCID; Morphology; Penicillin; Pluripotency; Proteins; RNA, Messenger - chemistry; RNA, Messenger - genetics; Secretion; Stem cells; Technology application; Therapeutic applications; Transcription factors; Transfection; α-Fetoprotein
• ISSN: 2314-6133; 2314-6141
• DOI: 10.1155/2018/8240567

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 F a h 1 R T y r c /RJ models. In conclusion, our data suggest that the nonintegrating method using mRNA has potential for cell therapy.