Expansion of murine spermatogonial stem cells through serial transplantation
Expansion of murine spermatogonial stem cells through serial transplantation
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DOI：
10.1095/biolreprod.102.004549
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发表时间：
2003-01-01
影响因子：
3.6
通讯作者：
Kubota, Y
中科院分区：
文献类型：
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作者：
Ogawa, T;Ohmura, M;Kubota, Y
哺乳动物雄性生殖细胞可以终生产生大量精子，因此被普遍认为具有无限的增殖潜力。然而，几乎没有实质性的证据支持这一假设。在本研究中，我们进行了连续移植的精原干细胞，以调查如果他们扩大自我更新分裂移植后。携带绿色荧光蛋白基因的转基因小鼠被用作供体细胞来源，其促进受体睾丸中定殖的供体生殖细胞的鉴定和回忆。收集受体睾丸中建立的生殖细胞集落并移植到新的受体中。这种精原干细胞的连续移植使受体睾丸重新增殖，从一个受体到下一个受体连续成功地进行了四次。两次连续移植之间的潜伏期为55至373天。在这些传代过程中，精原干细胞表现出恒定的活性，在受体睾丸中形成生精集落。在移植后的一年多里，他们的数量继续增加。以SI/SId小鼠为受体，仅在其睾丸中增殖未分化的A型精原细胞，精原干细胞的定殖效率为4.25%。基于定殖效率，一个集落形成活性被评估为等于约20个精原干细胞。在本实验中，精原干细胞估计在100天内扩增超过50倍。
Mammalian male germ cells might be generally thought to have infinite proliferative potential based on their life-long production of huge numbers of sperm. However, there has been little substantial evidence that supports this assumption. In the present study, we performed serial transplantation of spermatogonial stem cells to investigate if they expand by self-renewing division following transplantation. The transgenic mouse carrying the Green fluorescent protein gene was used as the donor cell source that facilitated identification and recollection of colonized donor germ cells in the recipient testes. The established colonies of germ cells in the recipient testes were collected and transplanted to new recipients. This serial transplantation of spermatogonial stem cells repopulated the recipient testes, which were successfully performed sequentially up to four times from one recipient to the next. The incubation periods between two sequential transplantations ranged from 55 to 373 days. During these passages, the spermatogonial stem cells showed constant activity to form spermatogenic colonies in the recipient testis. They continued to increase in number for more than a year following transplantation. Colonization efficiency of spermatogonial stem cells was determined to be 4.25% by using SI/SId mice as recipients that propagated only undifferentiated type A spermatogonia in their testes. Based on the colonization efficiency, one colony-forming activity was assessed to equate to about 20 spermatogonial stem cells. The spermatogonial stem cells were estimated to expand over 50-fold in 100 days in this experiment.