• Cytokines;
  • Spermatogenesis;
  • Spermatogonial stem cells;
  • Three-dimension system;
  • Pluripotent stem cells;
  • Stem cell-microenvironment


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. STEM CELLS2012;30:2355–2360