Analysis of spermatogonial proliferation with technique of spermatogonial transplantation-A trial for improving spermatogenic activity

精原细胞移植技术分析精原细胞增殖-提高生精活性的尝试

• 批准号: 11671569
• 负责人: OGAWA Takehiko
• 金额: $ 1.41万
• 依托单位: Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671569/
• 关键词: Spermatogeneiss; Spermatogonia; Transplantation; Stem cell; GFP; 男性不妊; GnRH; アンドロゲン

Spermatogonial transplantation has several unique advantages when applied to the study of spermatogenesis. For instance, germ cells and testis environment composed of many somatic cells can be differently treated to see effects of the treatments. The proliferation status of spermatogonia can be also clearly observed in the early stage of colony development after transplantation. We have first introduced GFP mice in this system as donor mice by which the system became more quantitative. With GFP mice as donor, sequential transplantation of spermatogonial stem cell became possible. We have so far succeeded to passage stem cells for 4 times successively. Accumulated data indicated that spermatogonial stem cells expand as colony size increases, and as time passes by. The expansion rate was estimated to be about 30-fold during 100 days following transplantation. This expansion rate appeared to be constant for more than a year showing no sign of exhaustion. Meanwhile, we also tried to find the effect of intra-testicular microenvironment on spermatogonial proliferation. The LH-RH analogue (leuprorelin) was used to modify hormonal environment of the testis. In the leuprorelin-treated mouse testes, transplanted speramtogonia presented more densely on the basement membrane of the seminiferous tubules as early as 4 weeks after the transplantation. This finding suggests that spermatogonia are stimulated to proliferate or their apoptosis are reduced in the leuprorelin-treated mouse testes. Our present research established the spermatogonial transplantation technique as a quantitative method for evaluating various treatments on spermatogonial proliferation, differentiation, and death.

精原细胞移植应用于精子发生研究时具有几个独特的优势。例如，可以对由许多体细胞组成的生殖细胞和睾丸环境进行不同的处理，以观察治疗的效果。在移植后集落发育的早期阶段也可以清楚地观察到精原细胞的增殖状况。我们首先在该系统中引入 GFP 小鼠作为供体小鼠，使系统变得更加定量。以GFP小鼠为供体，精原干细胞的顺序移植成为可能。迄今为止，我们已连续成功进行了4次干细胞传代。累积的数据表明，精原干细胞随着集落大小的增加和时间的推移而扩张。移植后 100 天内，扩增率估计约为 30 倍。这种扩张速度似乎在一年多的时间里一直保持不变，没有显示出耗尽的迹象。同时，我们也尝试寻找睾丸内微环境对精原细胞增殖的影响。 LH-RH 类似物（亮丙瑞林）用于改变睾丸的激素环境。在亮丙瑞林处理的小鼠睾丸中，移植的精原细胞早在移植后 4 周就在生精小管的基底膜上出现更密集。这一发现表明，在亮丙瑞林处理的小鼠睾丸中，精原细胞增殖受到刺激，或者细胞凋亡减少。我们目前的研究建立了精原细胞移植技术作为评估各种治疗方法对精原细胞增殖、分化和死亡的定量方法。

项目成果

Parreira GG, Ogawa T, Avarbock MR, Franca LR, Hausler CL, Brinster RL, Russell LD: "Development of germ cell transplants : morphometric and ultrastructural studies."Tissue Cell. 31. 242-254 (1999)

Parreira GG、Okawa T、Avarbock MR、Franca LR、Hausler CL、Brinster RL、Russell LD：“生殖细胞移植的发展：形态测量和超微结构研究。”组织细胞。

Dobrinski I et al.: "Computer assisted image analysis to assess colonization of recipient seminiferous tubules by spermatogonial stem cells from transgenic donor mice."Molecular Reproduction and Development . 53. 142-148 (1999)

Dobrinski I 等人：“计算机辅助图像分析来评估来自转基因供体小鼠的精原干细胞对受体生精小管的定植。”分子繁殖和发育。

Ogawa T et al.: "Recipient preparation is critical for spermatogonial transplantation in the rat."Tissue Cell. 31・5. 461-472 (1999)

小川 T 等人：“受体准备对于大鼠精原细胞移植至关重要”。《组织细胞》31·5（1999）。

Parreira GG et al.: "Development of germ cell transplants : morphometric and ultrastructural studies."Tissue Cell. 31・3. 242-254 (1999)

Parreira GG 等人：“生殖细胞移植的发展：形态测量和超微结构研究”。《组织细胞》31·3（1999）。

Ogawa T et al.: "Recipient preparation is critical for spermatogonial transplantation in the rat."Tissue Cell. 31. 461-472 (1999)

Okawa T 等人：“受体准备对于大鼠精原细胞移植至关重要。”组织细胞。