权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Molecular mechanism of proliteration and differentistion of male germ cells analyzed by using the cell line that undergoes "meiosis"in vitro.

利用体外“减数分裂”细胞系分析雄性生殖细胞增殖分化的分子机制。

基本信息

批准号：
14370511
负责人：
FUJITA Jun
金额：
$ 6.27万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370511/
关键词：
meiosis germ cell antibody testis ovary tumor 培養細胞モノクローナル抗体免疫組織染色

项目摘要

1. Using the GC-2M cell line, a clonal line derived from the mouse spermatocytic cell line GC-2spd(ts) and thought to undergo meiosis in vitro, we obtained the following results.By RT-PCR, no expression of genes known to be specific to meiosis was detected in GC-2M. Karyotype analysis demonstrated that most cells have 74-75 chromosomes, near tetraploid, with some cells containing around 37 chromosomes, and that no haploid cells were ~present. Flowcytomery showed that the major peak in GC-2M as well as NIH/3T3 cells corresponded to 4n DNA contend Clonal culture of GC-2M cells with 4n or 8n DNA content suggested that they could proliferate and reduce the DNA content to 2n. These results indicate that the reduction of the number of chromosomes occurs in GC-2M cells, but the process is different from regular meiosis.We raised mouse monoclonal antibodies against GC-2M. cell lysates, and obtained 10 clones. They immunohistochemically stained the sections of mouse testis. Trials to identify the proteins recognized by the antibodies were not successful.2. We have established the first human ovarian dysgerminoma cell line HDGO. The karyotype was 46, XX, and expression of Vasa, a germ cell specific, protein, was detected in the cells. HDGO cells formed tumors in SCID mice, and constitutive activation of c-Kit proto-oncogene was identified.

1.使用GC-2M细胞系，该细胞系源自小鼠精母细胞系GC-2spd(ts)并被认为在体外进行减数分裂，我们获得了以下结果。通过RT-PCR，在GC-2M中未检测到已知减数分裂特异性基因的表达。核型分析表明，大多数细胞有74-75条染色体，接近四倍体，一些细胞含有大约37条染色体，并且不存在单倍体细胞。流式细胞术显示GC-2M以及NIH/3T3细胞中的主峰对应于4n DNA含量。具有4n或8n DNA含量的GC-2M细胞的克隆培养表明它们可以增殖并将DNA含量降低至2n。这些结果表明GC-2M细胞中发生了染色体数量的减少，但该过程与常规减数分裂不同。我们制备了针对GC-2M的小鼠单克隆抗体。细胞裂解液，获得10个克隆。他们对小鼠睾丸切片进行了免疫组织化学染色。鉴定抗体识别的蛋白质的试验没有成功。2．我们建立了第一个人卵巢无性细胞瘤细胞系HDGO。核型为 46, XX，并在细胞中检测到 Vasa（一种生殖细胞特异性蛋白质）的表达。 HDGO 细胞在 SCID 小鼠中形成肿瘤，并鉴定出 c-Kit 原癌基因的组成型激活。