Functional analysis of spermatogonial stem cells by cell fusion

通过细胞融合对精原干细胞进行功能分析

• 批准号: 18590167
• 负责人: TAKEHASHI Masanori
• 金额: $ 2.55万
• 依托单位: Otani Womens University (2007)Kyoto University (2006)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590167/
• 关键词: spermatogonial stem cells; cell fusion; spermatogenesis; development and differentiation; genetics

Spermatogonial stem cells (SSCs) have unique properties to self-renew and support spermatogenesis throughout their lifespan. SSCs can proliferate in vitro in the presence of glial cell line-derived neurotrophic factor, a self-renewal factor for SSCs. These cultured SSCs are called germline stem (GS) cells. Although SSCs are normally committed to the germline, ES-like cell colonies develop spontaneously during GS cell culture from postnatal testis. These ES-like cells, called multipotent germline stem (mGS) cells, differentiate not only into somatic cells, but also into germ cells. In previous studies, it is known that germline cells such as ES cells and embryonic germ (EG) cells have the capacity to reprogram somatic nuclei after cell fusion. In this study, we investigated whether two type of germline cells derived from postnatal testis culture have the capacity similar to ES and EG cells. GS and mGS cells were established from newborn Green mice testes. These cells hybridized with thy … More mocytes, which derived from ROSA26 mice carrying a neo/lacZ transgene. It was possible to isolate hybrid clones by their resistance to G418 selection following cell fusion. GS-thymocyte hybrid cells were produced by electric or polyethylene glycol-mediated cell fusion ; however, these cells were not expanded by G418 selection because of low growth rate. mGS-thymocyte hybrid cells that maintained tetraploid chromosomes were clonally isolated by G418 selection and largely expanded. The morphology, growth rate, and characteristic gene expression patterns of these hybrid cells were similar to that of the mGS cells. These cells were able to differentiate into typical teratomas, which obtained tissues from three germ layers. Furthermore, the Oct-GFP transgene, which was repressed in thymocytes prior to cell fusion, was reactivated in the mGS-thymocyte hybrid cells. These results suggest that mGS cells through cell fusion can erase the developmental programming of somatic nulei and impose pluripotency. Less

精原干细胞（SSC）具有独特的自我更新特性，并在其整个生命周期中支持精子发生。SSCs可以在胶质细胞源性神经营养因子（一种SSCs的自我更新因子）的存在下在体外增殖。这些培养的SSC被称为生殖干细胞（GS）。虽然精原干细胞通常定向于生殖系，但ES样细胞集落在出生后睾丸的GS细胞培养过程中自发发育。这些ES样细胞，称为多能生殖干细胞（mGS），不仅分化成体细胞，而且分化成生殖细胞。在先前的研究中，已知生殖系细胞如ES细胞和胚胎生殖（EG）细胞具有在细胞融合后重编程体细胞核的能力。本研究旨在探讨两种来源于出生后睾丸培养的生殖系细胞是否具有类似于ES和EG细胞的能力。从新生绿色小鼠睾丸中建立GS和mGS细胞。这些细胞与 ...更多信息 mocytes，其来源于携带neo/lacZ转基因的ROSA 26小鼠。通过细胞融合后对G418选择的抗性，有可能分离杂种克隆。通过电融合或聚乙二醇介导的细胞融合产生GS-胸腺细胞杂交细胞;然而，由于生长速率低，这些细胞没有通过G418选择扩增。通过G418选择克隆分离保持四倍体染色体的mGS-胸腺细胞杂交细胞并大量扩增。这些杂交细胞的形态、生长速率和特征基因表达模式与mGS细胞相似。这些细胞能够分化成典型的畸胎瘤，其获得来自三个胚层的组织。此外，Oct-GFP转基因，这是抑制在胸腺细胞之前，细胞融合，被重新激活的mGS-胸腺细胞杂交细胞。这些结果表明，mGS细胞通过细胞融合可以消除体细胞核的发育编程，并施加多能性。少

项目成果

Rats produced by interspecies spermatogonial transplantation in mice and in vitro microinsemination

• DOI: 10.1073/pnas.0604205103
• 发表时间: 2006-09-12
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Shinohara, Takashi;Kato, Megumi;Hirabayashi, Masumi
• 通讯作者: Hirabayashi, Masumi

Production of knockout mice by gene targeting in multipotent germline stem cells.

• DOI: 10.1016/j.ydbio.2007.09.029
• 发表时间: 2007-12
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: M. Takehashi;Mito Kanatsu-Shinohara;H. Miki;Jiyoung Lee;Y. Kazuki;K. Inoue;N. Ogonuki;S. Toyokuni;M. Oshimura;A. Ogura;T. Shinohara

再生医療へ進む最先端の幹細胞研究(編集/山中伸弥、中内啓光:実験医学Vol.26 No.5増刊)

尖端干细胞研究向再生医学迈进（编辑/山中伸也、中内宏光：实验医学第26卷第5号特刊）

• 发表时间: 2008
• 作者: Yoshida;M.;Matzno;S.;Namba;H .;Nishikata;M.;Matsuyama;K.;竹橋 正則
• 通讯作者: 竹橋 正則

再生医療へ進む最先端の幹細胞研究(実験医学26巻5号(増刊))

尖端干细胞研究走向再生医学（实验医学第26卷第5期（特刊））

• 发表时间: 2008
• 作者: Sumio Matzno;Shinya Yasyda;Yuka Kitasa;Takeshi Akiyosi;Naoko Tanaka;Sachiko Juman;Kazumasa Shinozuka;Toshikatsu Nakabayasi;Kenji Matsuyama;竹橋正則
• 通讯作者: 竹橋正則

Clonal origin of germ cell colonies after spermatogonial transplantation in mice

• DOI: 10.1095/biolreprod.106.051193
• 发表时间: 2006-07-01
• 期刊: BIOLOGY OF REPRODUCTION
• 影响因子: 3.6
• 作者: Kanatsu-Shinohara, Mito;Inoue, Kimiko;Shinohara, Takashi
• 通讯作者: Shinohara, Takashi