Induced hepatic stem cells are suitable for human hepatocyte production

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

doi:10.1016/j.isci.2022.105052

Induced hepatic stem cells are suitable for human hepatocyte production

iScience. 2022 Sep 5;25(10):105052. doi: 10.1016/j.isci.2022.105052. eCollection 2022 Oct 21.

Authors

Yoshiki Nakashima^{1

2}, Chika Miyagi-Shiohira¹, Issei Saitoh³, Masami Watanabe⁴, Masayuki Matsushita⁵, Masayoshi Tsukahara², Hirofumi Noguchi¹

Affiliations

¹ Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, 207 Uehara, Nishihara, Okinawa 903-0215, Japan.
² Kyoto University Center for iPS Cell Research and Application Foundation (CiRA Foundation), Facility for iPS Cell Therapy (FiT), Kyoto 606-8397, Japan.
³ Division of Pediatric Dentistry, Graduate School of Medical and Dental Science, Niigata University, Niigata 951-8514, Japan.
⁴ Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan.
⁵ Department of Molecular and Cellular Physiology, Graduate School of Medicine, University of the Ryukyus, Okinawa 903-0215, Japan.

Abstract

Human hepatocytes were transfected with Sendai virus vectors (SeV) expressing OCT3/4, SOX2, KLF4, and C-MYC to produce hepatocyte-derived induced pluripotent stem cells (iPSCs). The messenger RNA (mRNA) expression of undifferentiated markers (passage 19-21) and hepatocyte-specific markers (HSMs) (passage 0-20) in 48 established hepatocyte-derived iPSC-like colonies was examined. Among the 48 clones, 10 clones continuously expressed HSM mRNA (HNF1β and HNF4α) in passage 0-20. The colonies which expressed HSMs (iTS-L cells: induced tissue-specific stem cells from liver) showed a different tendency in microarray and methylation analyses to fibroblast-derived iPSCs (strain: 201B7). iTS-L cells were less likely to form teratomas in mice than iPSCs (He). The iTS-L cells were differentiated into hepatocyte-like cells more efficiently than iPSCs (He) or iPSCs (201B7). These data suggest that SeV expressing OCT3/4, SOX2, KLF4, and C-MYC induce the generation of iPSCs and iTS-L cells.

Keywords: Human; biological sciences; cell biology; developmental biology; stem cells research.