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| 1 | Promoted differentiation of cynomolgus monkey ES cells into hepatocyte-like cells by co-culture with mouse fetal liver-derived cells显示文摘AIM: To explore whether a co-culture of cynomolgus monkey embryonic stem (cES) cells with embryonic liver cells could promote their differentiation into hepatocytes. METHODS: Mouse fetal liver-derived cells (MFLCs) were prepared as adherent cells from mouse embryos on embryonic d (ED) 14, after which undifferentiated cES cells were co-cultured with MFLCs. The induction of cES cells along a hepatic lineage was examined in MFLC- assisted differentiation, spontaneous differentiation, and growth factors (GF) and chemicals-induced differentiations (GF-induced differentiation) using retinoic acid, leukemia inhibitory factor (LIF), FGF2, FGF4, hepatocyte growth factor (HGF), oncostatin M (OSM), and dexamethasone. RESULTS: The mRNA expression of α-fetoprotein, albumin (ALB), α-1-antitrypsin, and hepatocyte nuclear factor 4α was observed earlier in the differentiating cES cells co-cultured with MFLCs, as compared to cES cells undergoing spontaneous differentiation and those subjected to GF-induced differentiation. The expression of cytochrome P450 7a1, a possible marker for embryonic endoderm-derived mature hepatocytes, was only observed in cES cells that had differentiated in a co-culture with MFLCs. Further, the disappearance of Oct3/4, a representative marker of an undifferentiated state, was noted in cells co-cultured with MFLCs, but not in those undergoing spontaneous or GF-induced differentiation. Immunocytochemical analysis revealed an increased ratio of ALB-immunopositive cells among cES cells co-cultured with MFLCs, while glycogen storageand urea synthesis were also demonstrated. CONCLUSION: MFLCs showed an ability to induce cES cells to differentiate toward hepatocytes. The co-culture system with MFLCs is a useful method for induction of hepatocyte-like cells from undifferentiated cES cells. | Ko Saito Masahide Yoshikawa Yukiteru Ouji Kei Moriya Mariko Nishiofuku Shigehiko Ueda Noriko Hayashi Shigeaki Ishizaka Hiroshi Fukui | 2006 | World Journal of Gastroenterology2006，12，42： | 11 |
| 2 | Differentiation of embryonic stem cells into insulin-producing cells promoted by Nkx2.2 gene transfer显示文摘AIM: To investigate the ability of a genetically altered embryonic stem (ES) cell line to generate insulin-producing cells in vitro following transfer of the Nkx2.2 gene.METHODS: Hamster Nkx2.2 genes were transferred into mouse ES cells. Parental and Nkx2.2-transfected ES cells were initiated toward differentiation in embryoid body (EB)culture for 5 d and the resulting EBs were transferred to an attached culture system. Dithizone (DTZ), a zincchelating agent known to selectively stain pancreatic beta cells, was used to detect insulin-producing cells.The outgrowths were incubated in DTZ solution (final concentration, 100 μg/mL) for 15 min before being examined microscopically. Gene expression of the endocrine pancreatic markers was also analyzed by RT-PCR. In addition, insulin production was determined immunohistochemically and its secretion was examined using an ELISA.RESULTS: DTZ-stained cellular clusters appeared after approximately 14 d in the culture of Nkx2.2-transfected ES cells (Nkx-ES cells), which was as much as 2 wk earlier, than those in the culture of parental ES cells (wt-ES). The frequency of DTZ-positive cells among total cultured cells on day 28 accounted for approximately 1.0% and 0.1% of the Nkx-ES- and wt-ES-derived EB outgrowths, respectively. The DTZ-positive cellular clusters were found to be immunoreactive to insulin, while the gene expressions of pancreatic-duodenal homeobox 1(PDX1), proinsulin 1 and proinsulin 2 were observed in the cultures that contained DTZ-positive cellular clusters.Insulin secretion was also confirmed by ELISA, whereas glucose-dependent secretion was not demonstrated.CONCLUSION: Nkx2.2-transfected ES cells showed an ability to differentiate into insulin-producing cells. | Akira Shiroi Shigehiko Ueda Yukiteru Ouji Ko Saito Kei Moriya Yuko Sugie Hiroshi Fukui Shigeaki Ishizaka Masahide Yoshikawa | 2005 | World Journal of Gastroenterology2005，11，27： | 9 |
| 3 | Embryonic stem cells develop into hepatocytes after intrasplenic transplantation in CCl_4-treated mice显示文摘AIM: To transplant undifferentiated embryonic stem (ES) cells into the spleens of carbon tetrachloride (CCl4)-treated mice to determine their ability to differentiate into hepatocytes in the liver. METHODS: CCl4, 0.5 mL/kg body weight, was injected into the peritoneum of C57BL/6 mice twice a week for 5 wk. In group 1 (n = 12), 1 x 105 undifferentiated ES cells (0.1 mL of 1 x 106/mL solution), genetically labeled with GFP, were transplanted into the spleens 1 d after the second injection. Group 2 mice (n = 12) were injected with 0.2 mL of saline twice a week, instead of CCl4, and the same amount of ES cells was transplanted into the spleens. Group 3 mice (n = 6) were treated with CCl4 and injected with 0.1 mL of saline into the spleen, instead of ES cells. Histochemical analyses of the livers were performed on post-transplantation d (PD) 10, 20, and 30. RESULTS: Considerable numbers of GFP-immunopositive cells were found in the periportal regions in group 1 mice (CCl4-treated) on PD 10, however, not in those untreated with CCl4 (group 2). The GFP-positive cells were also immunopositive for albumin (ALB), alpha-1 antitrypsin, cytokeratin 18, and hepatocyte nuclear factor 4 alpha on PD 20. Interestingly, most of the GFP-positive cells were immunopositive for DLK, a hepatoblast marker, on PD 10. Although very few ES-derived cells were demonstrated immunohistologically in the livers of group 1 mice on PD 30, improvements in liver fibrosis were observed. Unexpectedly, liver tumor formation was not observed in any of the mice that received ES cell transplantationduring the experimental period. CONCLUSION: Undifferentiated ES cells developed into hepatocyte-like cells with appropriate integration into tissue, without uncontrolled cell growth. | Kei Moriya Masahide Yoshikawa Ko Saito Yukiteru Ouji Mariko Nishiofuku Noriko Hayashi Shigeaki Ishizaka Hiroshi Fukui | 2007 | World Journal of Gastroenterology2007，13，6： | 5 |
| 4 | Wnt-10b promotes differentiation of skin epithelial cells in vitro显示文摘 | Ouji Y Yoshikawa M Shiroi A | 2006 | Biochem Biophys Res Commun2006，342，1： | 1 |
| 5 | Isolation and characterization of murine hepatocytes following collagenase infusion into left ventricle of heart显示文摘 | Ouji Y Yoshikawa M Moriya K | 2010 | J Biosci Bioeng2010，110，4： | 1 |
| 6 | Promotion of hair follicle devdopment and tfichogenesis by Wnt-10b in cul- tured embryonic skin and in reconstituted skin 显示文摘 | OUJI Y YOSHIKAWA M SHIROI A | 2006 | Biochem Biophys Res Commun2006，345，2： | 1 |
| 7 | Wnt-10b pro- motes differentiation of skin epithelial cells in vitro显示文摘 | OUJI Y YOSHIKAWA M SHIROI A | 2006 | Biochem Biophys Res Commun2006，342，1： | 1 |
| 8 | 显示文摘 | Kanda S Shiroi A Ouji Y | 2003 | Hepatol Res2003，26，3： | 1 |
| 9 | Wnt-10b, u- niqudy among Wnts, promotes epithelial differentiation and shaft growth显示文摘 | OUJI Y YOSHIKAWA M MOR1YA K | 2008 | Biochem Biophys Res Commun2008，367，2： | 1 |
| 10 | Promotion of hair follicle development and trichogenesis by Wnt-10b in cultured embryonic skin and in reconstituted skin显示文摘 | Ouji Y Yoshikawa M Shiroi A | | 0，，2： | 1 |
| 11 | Canonical Wnts,specifically Wnt-10b,show ability to maintain dermal papilla cells显示文摘 | Ouji Y Nakamura-Uchiyama F Yoshikawa M | 2013 | Biochem Biophys Res Commun2013，438，3： | 1 |
| 12 | Cytotoxic human peripheral blood-derived γδT cells kill glioblastoma cell lines: implications for cell-based immunotherapy for patients with glioblastoma显示文摘 | Tsutomu Nakazawa Mitsutoshi Nakamura Young Soo Park Yasushi Motoyama Yasuo Hironaka Fumihiko Nishimura Ichiro Nakagawa Shuichi Yamada Ryosuke Matsuda Kentaro Tamura Tadashi Sugimoto Yasuhiro Takeshima Akiko Marutani Takahiro Tsujimura Noriko Ouji Yukiteru | 2014 | Journal of Neuro-Oncology2014，，1： | 1 |
| 13 | Promotion ofhair follicle devel-opmentand trichogenesis by Wnt10b in cultured embryonic skin and in reconstituted skin显示文摘 | Ouji Y Yoshikawa M Shiroi A | 2006 | Biochem Biophys Res Commun2006，345，2： | 1 |
| 14 | Canonical Wnts, specifically Wnt-10b, show ability to maintain dermal papillacells 显示文摘 | Ouji Y Nakamura-Uehiyama F Yoshikawa M | 2013 | Biochem Biophys Res Commun2013，438，3： | 1 |
| 15 | Wnt-10b promotes dif- ferentiation of skin epithelial cells in vitro 显示文摘 | Ouji Y Yoshikawa M Shiroi A | 2006 | Biochem Biophys Res Comrnun2006，342，1： | 1 |
| 16 | Wnt-10b promotes differentiation of skin epithelial cells in vitro 显示文摘 | Ouji Y Yoshikawa M Shiroi A | 2006 | Biochemical and Biophysical Research Communications2006，3，1： | 1 |
| 17 | Promotion of hair follicle development and trichogenesis by Wnt-10b in cultured embryon- ic skin and in reconstituted skin显示文摘 | Ouji Y Yoshikawa M Shiroi A | 2006 | Biochem Biophys Res Com- mun2006，345，2： | 1 |
| 18 | Effects of Wnt-10b on hair shaft growth in hair follicle cultures显示文摘 | Ouji Y Yoshikawa M Shiroi A | 2007 | Biochem Biophys Res Com- mun2007，359，： | 1 |
| 19 | In vitro dif- ferentiation of mouse embryonic stem cells into inner ear hair cell-like cells using stromal cell conditioned medium显示文摘 | Ouji Y Ishiza S Nakamura-Uchiyama F | 2012 | Cell Death Dis2012，3，： | 1 |
| 20 | Wnt-10b promotes differentiation of skin epithelial cells in vitro 显示文摘 | Ouji Y Yoshikawa M Shiroi A | 2006 | Biochemical and Biophysical Res earch Communications2006，342，1： | 1 |
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