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

Hirofumi Noguchi; Yoshiki Nakashima; Masami Watanabe; Masayuki Matsushita; Masayoshi Tsukahara; Issei Saitoh; Chika Miyagi-Shiohira

doi:10.1016/j.xpro.2022.101884

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

Hirofumi Noguchi, Yoshiki Nakashima, Masami Watanabe, Masayuki Matsushita, Masayoshi Tsukahara, Issei Saitoh, Chika Miyagi-Shiohira

Research output: Contribution to journal › Article › peer-review

Abstract

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

Original language	English
Article number	101884
Journal	STAR Protocols
Volume	3
Issue number	4
DOIs	https://doi.org/10.1016/j.xpro.2022.101884
Publication status	Published - Dec 16 2022
Externally published	Yes

Keywords

Cell culture
Cell Differentiation
Health Sciences
Stem Cells

ASJC Scopus subject areas

Neuroscience(all)
Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)

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10.1016/j.xpro.2022.101884

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title = "Protocol for the generation of human induced hepatic stem cells using Sendai virus vectors",

abstract = "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",

keywords = "Cell culture, Cell Differentiation, Health Sciences, Stem Cells",

author = "Hirofumi Noguchi and Yoshiki Nakashima and Masami Watanabe and Masayuki Matsushita and Masayoshi Tsukahara and Issei Saitoh and Chika Miyagi-Shiohira",

note = "Funding Information: This work was supported in part by JSPS KAKENHI (grant numbers JP22K08759 , JP21K19537 , JP20H03745 , and JP19K09051 ), the Okinawa Science and Technology Innovation System Construction Project (OSTC), and the Japan Agency for Medical Research and Development (AMED) (grant number JP22bm0104001 ). Publisher Copyright: {\textcopyright} 2022 The Author(s)",

year = "2022",

month = dec,

day = "16",

doi = "10.1016/j.xpro.2022.101884",

language = "English",

volume = "3",

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TY - JOUR

T1 - Protocol for the generation of human induced hepatic stem cells using Sendai virus vectors

AU - Noguchi, Hirofumi

AU - Nakashima, Yoshiki

AU - Watanabe, Masami

AU - Matsushita, Masayuki

AU - Tsukahara, Masayoshi

AU - Saitoh, Issei

AU - Miyagi-Shiohira, Chika

N1 - Funding Information: This work was supported in part by JSPS KAKENHI (grant numbers JP22K08759 , JP21K19537 , JP20H03745 , and JP19K09051 ), the Okinawa Science and Technology Innovation System Construction Project (OSTC), and the Japan Agency for Medical Research and Development (AMED) (grant number JP22bm0104001 ). Publisher Copyright: © 2022 The Author(s)

PY - 2022/12/16

Y1 - 2022/12/16

N2 - 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

AB - 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

KW - Cell culture

KW - Cell Differentiation

KW - Health Sciences

KW - Stem Cells

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ER -

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

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Keywords

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