The Shraga Segal Department of Microbiology and Immunology, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
Stem Cells. 2012 Nov;30(11):2355-60. doi: 10.1002/stem.1238.
Culture of spermatogonial stem cells has been performed under a variety of conditions. Most featured two-dimensional systems, with different types of sera, conditioned media, feeder layers, and growth factors. Some have used three-dimensional (3D) matrices produced from gelatin, collagen, or other material. In spite of their increasingly sophisticated composition, however, complete spermatogenesis in vitro has not yet been achieved. In the seminiferous tubules, spermatogenesis occurs in an environment where cells are embedded in a 3D structure with specific niches regulating each stage of germ cell maturation mediated by hormones and paracrine/autocrine factors. We have recently reported achievement of complete in vitro spermatogenesis of mouse testicular germ cells in a 3D culture system featuring a soft agar matrix. This review discusses the advantages of the 3D culture system for studying the spermatogenic process in its entirety. Also discussed are the steps necessary to expand the applicability of the 3D culture system to human germ cell development and determine the functionality of culture-produced spermatozoa for generating offspring.
已经在多种条件下对精原干细胞进行了培养。大多数培养方法采用二维系统，使用不同类型的血清、条件培养基、饲养层和生长因子。有些方法使用了从明胶、胶原或其他材料制成的三维（3D）基质。然而，尽管它们的组成越来越复杂，但体外完全的精子发生尚未实现。在曲细精管中，精子发生发生在一个细胞嵌入具有特定小生境的 3D 结构的环境中，这些小生境通过激素和旁分泌/自分泌因子调节由生殖细胞成熟的各个阶段。我们最近报道了在具有软琼脂基质的 3D 培养系统中实现了小鼠睾丸生殖细胞的完全体外精子发生。本综述讨论了 3D 培养系统在研究整个精子发生过程中的优势。还讨论了将 3D 培养系统扩展应用于人类生殖细胞发育和确定培养产生的精子用于生育后代的功能的必要步骤。
