Use of 2D and 3D culture systems for differentiation of spermatogenic stem cells and induction of spermatogenesis in vitro in mouse

Abstract

The study of spermatogonial stem cells (SSC) and consequently of spermatogenesis, can provide a better model to clarify the biology of adult SC. Furthermore, understanding the mechanisms involved in functional control of the SSC may provide treatments for male infertility, understanding the origin and formation of testicular tumors, preservation of endangered species and genetic improvement for livestock, among other important issues in reproductive medicine. However there are few studies directly comparing cropping systems for the in vitro development of SSC. Thus, the general aim of this experiment is to study cell differentiation and induction of spermatogenesis in vitro in mice spermatogonial stem cells grown in 2D and 3D systems. To do so will be sacrificed mice 7 days old, who will have his testicles removed and macerdaos to separate the SSC. This will be grown in cell suspension system consisting of two: 2D - slabs of cultivation covered by a thin layer of jelly; 3D - soft agar matrix (SACS). Also will be studied in two cropping systems, the effect of the addition of gonadotropins to the culture medium in the success of meiotic progression of germ cells. At the beginning of cultivation, as well as their deferent stages will be characterized by immunocytochemistry, the cells developed during spermatogenesis in vitro, as well as somatic cells co-cultured. Also be quantified by propidium iodide staining, the load of nuclear DNA of cultured cells. (AU)