Hepatic differentiation of human bone marrow-derived mesenchymal stem cells by tetracycline-regulated hepatocyte nuclear factor 3beta.
Human bone marrow-derived mesenchymal stem cells (BM-MSCs) are expected to be a potential source of cells for transplantation. Although recent reports have shown that isolated MSCs can differentiate into hepatocytes, the efficiency of differentiation is insufficient for therapeutic application. To circumvent this problem, it is necessary to understand the mechanisms of hepatic differentiation of human BM-MSCs. Hepatocyte nuclear factor 3beta (HNF3beta), a forkhead/winged helix transcription factor, is essential for liver development. In the present study, we established a tetracycline (Tet)-regulated expression system for HNF3beta in UE7T-13 BM-MSCs. HNF3beta expression significantly enhanced expression of albumin, alpha-fetoprotein (AFP), tyrosine amino transferase (TAT) and epithelial cell adhesion molecule (EpCAM) genes. The differentiated cells showed hepatocyte-specific functions including glycogen production and urea secretion. During treatment with the Tet-on system for 8 days, over 80% of UE7T-13 cells turned out to express albumin. Furthermore, the combination of Tet with basic fibroblast growth factor (bFGF) efficiently induced the genes such as albumin and TAT, which are associated with maturity of hepatocytes; however, it suppressed genes such as AFP and EpCAM, which are associated with immaturity of hepatocytes, suggesting that Tet-induced HNF3beta expression sensitizes BM-MSCs to bFGF signals. Finally, the results of the present study suggest that down-regulation of Wnt/beta-catenin signals caused by translocation of beta-catenin to cytoplasmic membrane is associated with hepatic differentiation of human BM-MSCs.


CONCLUSIONS
HNF3beta expression induced efficient differentiation of UE7T-13 human BM-MSCs.
Ishii
Kyoko
K
Department of Genetic Medicine and Regenerative Therapeutics, Division of Molecular and Genetic Medicine, Graduate School of Medicine, Tottori University, Yonago, Japan.
Yoshida
Yoko
Y
Akechi
Yuji
Y
Sakabe
Tomohiko
T
Nishio
Ren
R
Ikeda
Remina
R
Terabayashi
Kei
K
Matsumi
Yoshiaki
Y
Gonda
Kazue
K
Okamoto
Hideharu
H
Takubo
Kazuko
K
Tajima
Fumihito
F
Tsuchiya
Hiroyuki
H
Hoshikawa
Yoshiko
Y
Kurimasa
Akihiro
A
Umezawa
Akihiro
A
Shiota
Goshi
G
D000418:Albumins / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D016207:Cytokines / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D006003:Glycogen / Q000096:biosynthesis; D051542:Hepatocyte Nuclear Factor 3-beta / Q000235:genetics / Q000378:metabolism*; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D013752:Tetracycline / Q000008:administration & dosage / Q000494:pharmacology*; D014162:Transfection; D014508:Urea / Q000378:metabolism; D051153:Wnt Proteins / Q000378:metabolism; D051176:beta Catenin / Q000378:metabolism
D000418:Albumins; D016207:Cytokines; D051153:Wnt Proteins; D051176:beta Catenin; D016222:Fibroblast Growth Factor 2; D051542:Hepatocyte Nuclear Factor 3-beta; D014508:Urea; D006003:Glycogen; D013752:Tetracycline