In Vitro Spermatogenesis in Explanted Adult Mouse Testis Tissues.
In Vitro Spermatogenesis in Explanted Adult Mouse Testis Tissues.
复制标题
DOI：
10.1371/journal.pone.0130171
复制
发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Ogawa T
中科院分区：
文献类型：
--
作者：
Sato T;Katagiri K;Kojima K;Komeya M;Yao M;Ogawa T
精子发生的体外研究对于阐明精子发生的机制具有重要意义。我们以前开发了一种器官培养方法，可以支持精子发生从精原干细胞到精子形成使用未成熟的小鼠睾丸组织。在这项研究中，我们研究了它是否也适用于成年小鼠的成熟睾丸组织。我们使用了两种转基因小鼠品系，顶体蛋白-GFP和Gsg 2-GFP，它们分别携带对减数分裂和单倍体细胞特异性的标记GFP基因。在补充有10%KSR或AlbuMAX的培养基中培养在完全延伸时表达GFP的4至29周龄的成年GFP小鼠的睾丸组织片段。GFP表达迅速下降，在培养的第7至14天达到最低，但在随后的培养期间略有增加。这种增加反映了从头精子发生，证实了BrdU标记精母细胞和精子细胞。我们还使用了维生素A缺乏的小鼠，其睾丸仅含有精原细胞。这些小鼠在13-21周龄时的睾丸在培养时没有GFP表达，在培养过程中获得GFP表达，并在组织学上证实了精子发生。此外，Sl/Sld突变小鼠的成年睾丸组织，由于Kit配体突变而缺乏精子发生，与重组Kit配体一起培养以诱导精子发生直至单倍体形成。虽然精子发生率低于幼鼠，但本研究结果表明，器官培养法是培养成熟成年小鼠睾丸组织的有效方法，证明了精子发生从精原细胞到单倍体细胞的诱导。
Research on in vitro spermatogenesis is important for elucidating the spermatogenic mechanism. We previously developed an organ culture method which can support spermatogenesis from spermatogonial stem cells up to sperm formation using immature mouse testis tissues. In this study, we examined whether it is also applicable to mature testis tissues of adult mice. We used two lines of transgenic mice, Acrosin-GFP and Gsg2-GFP, which carry the marker GFP gene specific for meiotic and haploid cells, respectively. Testis tissue fragments of adult GFP mice, aged from 4 to 29 weeks old, which express GFP at full extension, were cultured in medium supplemented with 10% KSR or AlbuMAX. GFP expression decreased rapidly and became the lowest at 7 to 14 days of culture, but then slightly increased during the following culture period. This increase reflected de novo spermatogenesis, confirmed by BrdU labeling in spermatocytes and spermatids. We also used vitamin A-deficient mice, whose testes contain only spermatogonia. The testes of those mice at 13-21 weeks old, showing no GFP expression at explantation, gained GFP expression during culturing, and spermatogenesis was confirmed histologically. In addition, the adult testis tissues of Sl/Sld mutant mice, which lack spermatogenesis due to Kit ligand mutation, were cultured with recombinant Kit ligand to induce spermatogenesis up to haploid formation. Although the efficiency of spermatogenesis was lower than that of pup, present results showed that the organ culture method is effective for the culturing of mature adult mouse testis tissue, demonstrated by the induction of spermatogenesis from spermatogonia to haploid cells.
登录
查看更多内容
影响因子：
3.8
作者：
Schlatt, S;Kim, SS;Gosden, R
通讯作者：
Gosden, R
影响因子：
2.6
作者：
Ventel채, S;Mulari, M;Parvinen, M
通讯作者：
Parvinen, M
影响因子：
64.8
作者：
Honaramooz, A;Snedaker, A;Schlatt, S
通讯作者：
Schlatt, S
影响因子：
2.6
作者：
Sugimoto, Ryo;Nabeshima, Yo-ichi;Yoshida, Shosei
通讯作者：
Yoshida, Shosei
影响因子：
6.5
作者：
SUMMERLIN, WT;CHARLTON, E;KARASEK, M
通讯作者：
KARASEK, M