Ex vivo development of the entire mouse fetal reproductive tract by using microdissection and membrane-based organ culture techniques.
Ex vivo development of the entire mouse fetal reproductive tract by using microdissection and membrane-based organ culture techniques.
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DOI：
10.1016/j.diff.2022.01.001
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发表时间：
2022-01
期刊：
影响因子：
--
通讯作者：
Zhao F
中科院分区：
文献类型：
--
作者：
Jia S;Zhao F
离体外植体培养是胎儿生殖器官发育体内研究的一个有吸引力的替代方案。关于胎儿性腺的体外培养方法有大量文献。然而，尚未记录培养具有不同形状和大小的整个胎儿生殖道的方法。在这里，通过仔细的解剖和正确的组织方向，我们成功地在 Transwell 插入膜上培养了来自两性小鼠胚胎的整个双角生殖道。培养的生殖道系统经历了两性二态性的建立和区域特异性的形态发生，与体内生殖道系统的对应物的发育相当。为了测试这种培养方法的应用，我们使用化学处理（二氢睾酮和 BMS 564929）和遗传细胞消融小鼠模型（Gli1-CreER；Rosa-DTA）来研究雄激素信号传导和 Gli1+ 间充质在沃尔夫管发育中的作用。二氢睾酮和 BMS 564929 促进培养的 XX 组织中沃尔夫管的异位维持。在培养的组织中成功实现了 Gli1+ 间充质的高效和特异性消除，导致沃尔夫管卷曲缺陷。这些结果证明了这种器官培养方法适用于化学和遗传操作，否则很难在体内研究。总而言之，这种器官培养方法的建立为了解小鼠胎儿生殖道发育的体内研究提供了宝贵的补充工具。
Ex vivo explant culture is an appealing alternative to in vivo studies on fetal reproductive organ development. There is extensive literature on ex vivo methods of growing the fetal gonad. However, a method for culturing the whole fetal reproductive tract that has a different shape and size has not been documented. Here, with careful dissection and proper tissue orientation, we successfully cultured the entire bicornuate reproductive tracts from mouse embryos of both sexes on the Transwell insert membrane. The cultured reproductive tract system undergoes sexually dimorphic establishment and region-specific morphogenesis comparable to in vivo development of their counterparts. To test this culture method’s applications, we used chemical treatment (dihydrotestosterone and BMS 564929) and genetic cellular ablation mouse model (Gli1-CreER; Rosa-DTA) to investigate the roles of androgen signaling and Gli1+ mesenchyme in Wolffian duct development. The dihydrotestosterone and BMS 564929 promoted the ectopic maintenance of Wolffian ducts in cultured XX tissues. The efficient and specific elimination of Gli1+ mesenchyme was successfully achieved in the cultured tissues, resulting in defective coiling of Wolffian ducts. These results demonstrate the amenability of this organ culture method for chemical and genetic manipulations that are otherwise difficult to study in vivo. Taken together, the establishment of this organ culture method provides a valuable tool complementary to in vivo studies for understanding fetal reproductive tract development in mice.
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DOI：
10.1016/j.diff.2020.11.004
发表时间：
2021-03
期刊：
Differentiation; research in biological diversity
影响因子：
--
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通讯作者：
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