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

STAR Protoc. 2022 Dec 16;3(4):101884. doi: 10.1016/j.xpro.2022.101884. Epub 2022 Nov 23.

Authors

Hirofumi Noguchi¹, Yoshiki Nakashima², Masami Watanabe³, Masayuki Matsushita⁴, Masayoshi Tsukahara⁵, Issei Saitoh⁶, Chika Miyagi-Shiohira⁷

Affiliations

¹ Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, Okinawa 903-0215, Japan. Electronic address: noguchih@med.u-ryukyu.ac.jp.
² Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, Okinawa 903-0215, Japan; Kyoto University Center for iPS Cell Research and Application Foundation (CiRA Foundation), Facility for iPS Cell Therapy (FiT), Kyoto 606-8397, Japan.
³ Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan.
⁴ Department of Molecular and Cellular Physiology, Graduate School of Medicine, University of the Ryukyus, Okinawa 903-0215, Japan.
⁵ Kyoto University Center for iPS Cell Research and Application Foundation (CiRA Foundation), Facility for iPS Cell Therapy (FiT), Kyoto 606-8397, Japan.
⁶ Division of Pediatric Dentistry, Graduate School of Medical and Dental Science, Niigata University, Niigata 951-8514, Japan.
⁷ Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, Okinawa 903-0215, Japan.

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 Differentiation; Cell culture; Health Sciences; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cellular Reprogramming*
Hepatocytes
Humans
Induced Pluripotent Stem Cells*
Kruppel-Like Factor 4
Sendai virus / genetics

Substances

Kruppel-Like Factor 4