TY - JOUR
T1 - Induced hepatic stem cells are suitable for human hepatocyte production
AU - Nakashima, Yoshiki
AU - Miyagi-Shiohira, Chika
AU - Saitoh, Issei
AU - Watanabe, Masami
AU - Matsushita, Masayuki
AU - Tsukahara, Masayoshi
AU - Noguchi, Hirofumi
N1 - Funding Information:
We thank Naomi Kakazu (University of the Ryukyus) for clerical assistance and Maki Higa, Yuki Kawahira, Saori Adaniya, and Nagisa Higa (University of the Ryukyus) for providing technical support. This work was supported by the Research Laboratory Center, Faculty of Medicine, and the Institute for Animal Experiments, Faculty of Medicine, University of the Ryukyus. 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). Author roles: study design, YN, CS, HN; conducted the study, YN, CS, HN; data collection, YN, CS, HN; data analysis, YN, CS, HN; data interpretation, YN, CS, HN; provided materials, IS, MW, MM, MT; drafted the article, YN, HN; revised the content of the article, YN, HN; approval of the final version of the article, YN, CS, IS, MW, MM, MT, HN. YN takes responsibility for the integrity of all of the data analyses. The authors declare no competing interests. We worked to ensure diversity in experimental samples through the selection of genomic datasets.
Funding Information:
We thank Naomi Kakazu (University of the Ryukyus) for clerical assistance and Maki Higa, Yuki Kawahira, Saori Adaniya, and Nagisa Higa (University of the Ryukyus) for providing technical support. This work was supported by the Research Laboratory Center , Faculty of Medicine , and the Institute for Animal Experiments , Faculty of Medicine , University of the Ryukyus . 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 Authors
PY - 2022/10/21
Y1 - 2022/10/21
N2 - 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.
AB - 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.
KW - biological sciences
KW - cell biology
KW - developmental biology
KW - Human
KW - stem cells research
UR - http://www.scopus.com/inward/record.url?scp=85138104134&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85138104134&partnerID=8YFLogxK
U2 - 10.1016/j.isci.2022.105052
DO - 10.1016/j.isci.2022.105052
M3 - Article
AN - SCOPUS:85138104134
SN - 2589-0042
VL - 25
JO - iScience
JF - iScience
IS - 10
M1 - 105052
ER -