Protocol for the generation of human induced hepatic stem cells using Sendai virus vectors

Our recent study demonstrated the generation of induced tissue-specific stem/progenitor (iTS/iTP) cells by the transient overexpression of reprogramming factors combined with tissue-specific selection. Here, we present a protocol to reprogram human hepatocytes to generate human induced tissue-specif...

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Autores principales: Noguchi, Hirofumi, Nakashima, Yoshiki, Watanabe, Masami, Matsushita, Masayuki, Tsukahara, Masayoshi, Saitoh, Issei, Miyagi-Shiohira, Chika
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700318/
https://www.ncbi.nlm.nih.gov/pubmed/36595898
http://dx.doi.org/10.1016/j.xpro.2022.101884
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author Noguchi, Hirofumi
Nakashima, Yoshiki
Watanabe, Masami
Matsushita, Masayuki
Tsukahara, Masayoshi
Saitoh, Issei
Miyagi-Shiohira, Chika
author_facet Noguchi, Hirofumi
Nakashima, Yoshiki
Watanabe, Masami
Matsushita, Masayuki
Tsukahara, Masayoshi
Saitoh, Issei
Miyagi-Shiohira, Chika
author_sort Noguchi, Hirofumi
collection PubMed
description Our recent study demonstrated the generation of induced tissue-specific stem/progenitor (iTS/iTP) cells by the transient overexpression of reprogramming factors combined with tissue-specific selection. Here, we present a protocol to reprogram human hepatocytes to generate human induced tissue-specific liver stem (iTS-L) cells. Human hepatocytes are transfected with Sendai virus vectors (SeV) expressing OCT3/4, SOX2, KLF4, and c-MYC. iTS-L cells continuously express mRNA of hepatocyte-specific markers (HNF1β and HNF4α) and do not form teratomas. For complete details on the use and execution of this protocol, please refer to Nakashima et al. (2022).(1)
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spelling pubmed-97003182022-11-27 Protocol for the generation of human induced hepatic stem cells using Sendai virus vectors Noguchi, Hirofumi Nakashima, Yoshiki Watanabe, Masami Matsushita, Masayuki Tsukahara, Masayoshi Saitoh, Issei Miyagi-Shiohira, Chika STAR Protoc Protocol Our recent study demonstrated the generation of induced tissue-specific stem/progenitor (iTS/iTP) cells by the transient overexpression of reprogramming factors combined with tissue-specific selection. Here, we present a protocol to reprogram human hepatocytes to generate human induced tissue-specific liver stem (iTS-L) cells. Human hepatocytes are transfected with Sendai virus vectors (SeV) expressing OCT3/4, SOX2, KLF4, and c-MYC. iTS-L cells continuously express mRNA of hepatocyte-specific markers (HNF1β and HNF4α) and do not form teratomas. For complete details on the use and execution of this protocol, please refer to Nakashima et al. (2022).(1) Elsevier 2022-11-23 /pmc/articles/PMC9700318/ /pubmed/36595898 http://dx.doi.org/10.1016/j.xpro.2022.101884 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Noguchi, Hirofumi
Nakashima, Yoshiki
Watanabe, Masami
Matsushita, Masayuki
Tsukahara, Masayoshi
Saitoh, Issei
Miyagi-Shiohira, Chika
Protocol for the generation of human induced hepatic stem cells using Sendai virus vectors
title Protocol for the generation of human induced hepatic stem cells using Sendai virus vectors
title_full Protocol for the generation of human induced hepatic stem cells using Sendai virus vectors
title_fullStr Protocol for the generation of human induced hepatic stem cells using Sendai virus vectors
title_full_unstemmed Protocol for the generation of human induced hepatic stem cells using Sendai virus vectors
title_short Protocol for the generation of human induced hepatic stem cells using Sendai virus vectors
title_sort protocol for the generation of human induced hepatic stem cells using sendai virus vectors
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700318/
https://www.ncbi.nlm.nih.gov/pubmed/36595898
http://dx.doi.org/10.1016/j.xpro.2022.101884
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