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.

当将精子移植应用于精子发生研究时，具有几个独特的优势。例如，通过许多体细胞组成的生殖细胞和睾丸环境可以有所不同，以查看治疗的影响。在移植后的菌落发育的早期发育的早期，还可以清楚地观察到精子的增殖状态。我们首先在该系统中引入了GFP小鼠，作为供体小鼠，通过该小鼠，该系统变得更加定量。用GFP小鼠作为供体，精子干细胞的顺序移植成为可能。到目前为止，我们已经成功地传递了4次。累积数据表明，随着菌落大小的增加，精子干细胞会扩展，并且随着时间的流逝。在移植后100天内估计膨胀率约为30倍。这种膨胀率似乎是恒定的一年以上，没有疲惫的迹象。同时，我们还试图找到农内微环境对精子增生的影响。 LH-RH类似物（Leuprorelin）用于修饰睾丸的激素环境。在亮纹蛋白治疗的小鼠睾丸中，移植的蜘蛛症早在移植后4周就在精液小管的地下膜上更密集。这一发现表明，刺激了精子症以增殖或在luprorelin治疗的小鼠睾丸中降低其凋亡。我们目前的研究确立了精子移植技术，作为一种定量方法，用于评估有关精子增生，分化和死亡的各种治疗方法。

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）。

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。

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）。