Preparation of enriched mouse syncitia-free pachytene spermatocyte cell suspensions.
Preparation of enriched mouse syncitia-free pachytene spermatocyte cell suspensions.
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DOI：
10.1007/978-1-61779-436-0_6
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发表时间：
2012
影响因子：
--
通讯作者：
Yan Wang;M. Culty
中科院分区：
文献类型：
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作者：
Yan Wang;M. Culty
精子发生包括有丝分裂、减数分裂和分化的一系列复杂步骤，从二倍体精原干细胞的定向分化开始，到单倍体精子的形成结束。啮齿动物模型已被常规用于研究生殖细胞发育和生殖毒理学，因为它们与人类模型有许多相似之处，同时具有在几周内重现通常需要数年时间才能完成的过程的优势。本文介绍了一种方法，分离亚群的成年生殖细胞，更具体地说，粗线期精母细胞，使用两个连续的梯度，而不使用淘洗离心机，一个专门的设备，在许多实验室不可用。此外，该方法的目的是分离富集粗线期精母细胞制剂缺乏污染合胞体，通常形成的有毒物质或环境的侮辱。
Spermatogenesis comprises a complex succession of steps of mitosis, meiosis, and differentiation, starting with the commitment of diploid spermatogonial stem cells to differentiate and ending with the formation of haploid spermatozoa. Rodent models have been routinely used to study germ cell development and reproductive toxicology, since they present many similarities with their human counterpart while offering the advantage of recapitulating within a few weeks a process that normally takes years to occur. This article describes a method to isolate subpopulations of adult germ cells, more specifically pachytene spermatocytes, using two successive gradients, without using an elutriation centrifuge, a specialized device not available in many laboratories. Moreover, the method was designed to isolate enriched pachytene spermatocytes preparations devoid of contaminating syncitia, often formed in response to toxicants or environmental insults.