著者

Ryoji KANEGI Shingo HATOYA Kazuto KIMURA Kyohei YODOE Toshiya NISHIMURA Kikuya SUGIURA Noritoshi KAWATE Toshio INABA

出版者

The Society for Reproduction and Development

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

pp.2022-038, (Released:2023-10-26)

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Induced pluripotent stem (iPS) cells are generated from somatic cells and can differentiate into various cell types. Therefore, these cells are expected to be a powerful tool for modeling diseases and transplantation therapy. Generation of domestic cat iPS cells depending on leukemia inhibitory factor has been reported; however, this strategy may not be optimized. Considering that domestic cats are excellent models for studying spontaneous diseases, iPS cell generation is crucial. In this study, we aimed to derive iPS cells from cat embryonic fibroblasts retrovirally transfected with mouse Oct3/4, Klf4, Sox2, and c-Myc. After transfection, embryonic fibroblasts were reseeded onto inactivated SNL 76/7 and cultured in a medium supplemented with basic fibroblast growth factor. Flat, compact, primary colonies resembling human iPS colonies were observed. Additionally, primary colonies were more frequently observed in the KnockOut Serum Replacement medium than in the fetal bovine serum (FBS) medium. However, enhanced maintenance and proliferation of iPS-like cells occurred in the FBS medium. These iPS-like cells expressed embryonic stem cell markers, had normal karyotypes, proliferated beyond 45 passages, and differentiated into all three germ layers in vitro. Notably, expression of exogenous Oct3/4, Klf4, and Sox2 was silenced in these cells. However, the iPS-like cells failed to form teratomas. In conclusion, this is the first study to establish and characterize cat iPS-like cells, which can differentiate into different cell types depending on the basic fibroblast growth factor.

著者

Takahiro USHIGUSA Yoshiyuki KOYAMA Tomoko ITO Kenichi WATANABE James K. CHAMBERS Aya HASEGAWA Kazuyuki UCHIDA Ryoji KANEGI Shingo HATOYA Toshio INABA Kikuya SUGIURA

出版者

JAPANESE SOCIETY OF VETERINARY SCIENCE

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.17-0466, (Released:2018-01-01)

被引用文献数

6

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By using a complex of DNA, polyethylenimine and chondroitin sulfate, the in vivo transfection of early secretory antigenic target-6 (ESAT-6) gene into tumor cells was found to cause significant suppression of the tumor growth. In order to apply the method in clinical cancer treatment in dogs and cats, mechanisms underlying the suppressive effects were investigated in a tumor-bearing mouse model. The transfection efficiency was only about 10%, but the transfection of ESAT-6 DNA nevertheless induced systemic immune responses against ESAT-6. By triple injection of ESAT-6 DNA at three day intervals, the tumor was significantly reduced and almost disappeared by 5 days after the start of treatment, and did not increase for more than 15 days after the final treatment. In the immunohistochemistry, a larger number of dendritic cells (DCs)/macrophages expressing ionized calcium-binding adapter molecule 1 and CD3+ T cells was observed in tumors treated with ESAT-6 DNA, and their population further increased significantly by day 5. Moreover, the amount of tumor necrosis factor, which is an apoptosis-inducing factor produced mainly by DCs/macrophages, was greater in the ESAT-6 DNA treated tumors than in controls, and increased with repeat of the treatment. These results indicate that in vivo transfection of ESAT-6 DNA into tumor cells elicits significant inhibition of tumor growth, by inducing potent activity of innate immunity mediated by DCs/macrophages, which may be followed by adaptive immunity against tumor associated antigens, which was elicited by the costimulation with ESAT-6 antigen.

著者

Hiromichi TAMADA Nahoko ADACHI Noritoshi KAWATE Toshio INABA Shingo HATOYA Tsutomu SAWADA

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.15-0520, (Released:2015-11-22)

被引用文献数

2

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Factors involved in patency of uterine cervices in the bitch with pyometra remain to be clarified. This study examined relationship between patency and mRNA levels for inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-1, COX-2 and prostaglandin E synthase (PGES) in the uterine cervix of bitches with pyometra. Cervical patency was measured by inserting the stainless steel rods with different diameter into cervical canals. Levels of mRNA expression were determined by semi-quantitative reverse transcription-polymerase chain reaction. The cervical patency was positively correlated with mRNA levels for COX-2 and PGES, but not those for iNOS and COX-1. The results suggest that gene expression of COX-2 and PGES may be involved in the regulation of patency in the uterine cervix of bitches with pyometra.

著者

Hidetaka NISHIDA Hiroshi TANAKA Masahiko KITAMURA Shingo HATOYA2 Kikuya SUGIURA2 Toshio INABA2 Masanari NAKAYAMA

出版者

JAPANESE SOCIETY OF VETERINARY SCIENCE

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.1203280802, (Released:2012-04-02)

被引用文献数

3 9

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Progressive ataxia and paralysis in three Miniature Dachshunds were found to be caused by idiopathic sterile pyogranulomatous inflammation of epidural fat between T5 and L4. All dogs were managed by hemilaminectomy and removal of epidural compressive material. Surgical findings and histopathological evaluation were necessary to diagnose epidural pyogranulomatous inflammation. A dog did not regain motor and sensor function after the surgery. Two dogs had exhibited improved neurological function after the surgery, but they recurred. Oral cyclosporine treatment was useful for their long remission. Idiopathic sterile pyogranulomatous inflammation of epidural fat can be considered to be a cause of thoracolumbar myelopathy in dogs.

著者

Takumi YOSHIDA Md Emtiaj ALAM Keisuke HANAFUSA Yasunori TSUJIMOTO Masaya TSUKAMOTO Ryoji KANEGI Toshio INABA Kikuya SUGIURA Shingo HATOYA

出版者

The Society for Reproduction and Development

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

pp.2021-084, (Released:2022-01-07)

被引用文献数

2

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We examined the effectiveness of saline, Euro-Collins solution (EC), and ET-Kyoto solution (ET-K) as preservation media for the cold storage of feline ovaries. Ovaries were maintained in these media at 4°C for 24, 48, or 72 h until oocyte retrieval. The ET-K group exhibited a higher oocyte maturation rate than the saline group after 72 h of storage. Moreover, ET-K could sustain the competence of the feline oocytes to cleave after 48 h, and the morula formation rate of the ET-K group was higher than that of the other groups after 24 and 48 h. Furthermore, the ET-K group exhibited a higher blastocyst formation rate than the other groups after storage for 24 h, and only ET-K retained the developmental competence in blastocysts after 48 h of storage. In addition, regarding the cell numbers of the blastocysts, there was no significant difference among the tested groups. In conclusion, our results indicate that ET-K is a suitable preservation medium for feline ovaries.

著者

Michi NAKAMURA Hidetaka NISHIDA Karin YOSHIZAKI Hideo AKIYOSHI Shingo HATOYA Kikuya SUGIURA Toshio INABA

出版者

JAPANESE SOCIETY OF VETERINARY SCIENCE

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.19-0141, (Released:2020-04-06)

被引用文献数

8

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We examined the paracrine action of canine mesenchymal stromal cells (MSCs) derived from bone marrow on the survival and differentiation of neural stem cells (NSCs) in vitro. MSCs were collected from the proximal end of the diaphysis of femur of healthy beagle dogs. The 70–80% confluent MSCs were re-fed with serum-free DMEM. The MSCs were incubated for 48 hr and the supernatant was collected as the conditioned medium (MSC-CM). The survival rate of NSCs in MSC-CM was significantly greater than in the medium without MSC-CM. The percentage of differentiated neurons and neurite length in MSC-CM was also significantly higher than in the medium without MSC-CM. These results suggested that canine MSC-CM promotes stem cell survival and neural differentiation of NSCs.

著者

Hiromichi TAMADA Kohei TAKEMOTO Masato TOMINAGA Noritoshi KAWATE Masahiro TAKAHASHI Shingo HATOYA Satoshi MATSUYAMA Toshio INABA Tsutomu SAWADA

出版者

日本繁殖生物学会

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

pp.2015-079, (Released:2015-10-26)

被引用文献数

2 4

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Gene expression of epidermal growth factor (EGF), transforming growth factor-α (TGF-α) and EGF receptor (EGF-R) and the localization of the corresponding proteins in the canine testis were studied. Levels of mRNA expressions were determined by semiquantitative reverse transcription polymerase chain reaction in the testes of the peripubertal (4–6 mo), young adult (3–4 y), advanced adult (7–8 y) and senescent (11–16 y) groups. The EGF-R mRNA level in the testes of the peripubertal group was significantly higher than those in the other groups, whereas there was no difference in EGF and TGF-α mRNA levels among groups. Immunohistochemical stainings for EGF, TGF-α and EGF-R in the testis revealed that immunoreactivity in the seminiferous epithelium and Sertoli cell was weak and nonspecific for the stage of spermatogenesis, and distinct staining was found in Leydig cells. These results suggest that the EGF family of growth factors may be involved in testicular maturation and function in the dog.

著者

Viskam WIJEWARDANA Kikuya SUGIURA Daluthgamage Patsy H. WIJESEKERA Shingo HATOYA Toshiya NISHIMURA Ryoji KANEGI Takahiro USHIGUSA Toshio INABA

出版者

公益社団法人 日本獣医学会

雑誌

Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

pp.14-0558, (Released:2015-02-20)

被引用文献数

1 6

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Previously, we reported that ovarian hormones affect the immune response against E. coli isolated from the dogs affected with pyometra. In order to investigate mechanisms underlying the immune modulation, we examined the effects of ovarian hormones on the generation of dendritic cells (DCs), the most potent antigen presenting cell. DCs were differentiated from peripheral blood monocytes (PBMOs) using a cytokine cocktail. Both estrogen receptor and progesterone receptors were expressed by the PBMOs and immature DCs. When various ovarian hormones were added to the culture for the DC differentiation, progesterone significantly decreased the expression of DC maturation markers, such as CD1a, CD80 and CD86, on mature DCs. Conversely, the addition of estrogen to the cultures increased the expression of CD86, but not other maturation makers. Furthermore, DCs differentiated in the presence of progesterone did not stimulate allogeneic mononuclear cells in PB. Taken together, these results indicate that progesterone diminishes the maturation of DCs, leading to decreased immune responses against invading pathogens.