Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cellsopen access
- Authors
- 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
- Issue Date
- Sep-2018
- Publisher
- HINDAWI LTD
- Citation
- BIOMED RESEARCH INTERNATIONAL, v.2018, pp.1 - 8
- Indexed
- SCIE
SCOPUS
- Journal Title
- BIOMED RESEARCH INTERNATIONAL
- Volume
- 2018
- Start Page
- 1
- End Page
- 8
- URI
- https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/16092
- DOI
- 10.1155/2018/8240567
- ISSN
- 2314-6133
- 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 alpha 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-iHceps expressed hepatocyte- specific marker genes and proteins, including albumin, alpha-fetoprotein, HNF4 alpha, 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 Fah(1R)Tyr(c)/RJ models. In conclusion, our data suggest that the nonintegrating method using mRNA has potential for cell therapy.
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