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Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cells
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 int...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6171260/ https://www.ncbi.nlm.nih.gov/pubmed/30327781 http://dx.doi.org/10.1155/2018/8240567 |
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author | 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 |
author_facet | 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 |
author_sort | Yoon, Sangtae |
collection | PubMed |
description | 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 Fah(1R)Tyr(c)/RJ models. In conclusion, our data suggest that the nonintegrating method using mRNA has potential for cell therapy. |
format | Online Article Text |
id | pubmed-6171260 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Hindawi |
record_format | MEDLINE/PubMed |
spelling | pubmed-61712602018-10-16 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 Biomed Res Int Research Article 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 Fah(1R)Tyr(c)/RJ models. In conclusion, our data suggest that the nonintegrating method using mRNA has potential for cell therapy. Hindawi 2018-09-20 /pmc/articles/PMC6171260/ /pubmed/30327781 http://dx.doi.org/10.1155/2018/8240567 Text en Copyright © 2018 Sangtae Yoon et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article 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 |
title | Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cells |
title_full | Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cells |
title_fullStr | Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cells |
title_full_unstemmed | Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cells |
title_short | Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cells |
title_sort | nonintegrating direct conversion using mrna into hepatocyte-like cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6171260/ https://www.ncbi.nlm.nih.gov/pubmed/30327781 http://dx.doi.org/10.1155/2018/8240567 |
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