Culture of mouse spermatogonial stem cells

Abstract

Spermatogenesis occurs within the seminiferous tubules of mammals by a complex process that is highly organized, extremely efficient and very productive. At the foundation of this process is the spermatogonial stem cell that is capable of both self-renewal and production of progeny cells, which undergo differentiation over a period of weeks to months in order to generate mature spermatozoa. It had been thought that germ cells survive only a brief period in culture, generally less than a few weeks. However, an accurate assessment of the presence of spermatogonial stem cells in any cell population has only recently become possible with development of the spermatogonial transplantation technique. Using this technique, we have demonstrated that mouse spermatogonial stem cells can be maintained in culture for approximately 4 months and will generate spermatogenesis following transplantation to the seminiferous tubules of an appropriate recipient. Extensive areas of cultured donor cell-derived spermatogenesis are generated in the host, and production of mature spermatozoa occurs. Cultivation of the testis cells on STO feeders is beneficial to stem cell survival. These results provide the first step in establishing a system that will permit spermatogonial stem cells to be cultivated and their number increased in vitro to allow for genetic modification before transplantation to a recipient testis.

Epigenetics and the plasticity of differentiation in normal and cancer stem cells

“…Epigenetic control of differentiation and transcription accessibility for genes of different tissues in normal adult stem cells Normal adult tissues contain stem cells that are responsible for regenerating these tissues (Spangrude et al, 1991;Nagano et al, 1998;Gage, 2000;Temple, 2001;Marshman et al, 2002;Alonso and Fuchs, 2003;Kubota et al, 2004;Collins et al, 2005;Shackelton et al, 2006). Adult stem cells reside in specific niches within different issues (Fuchs and Segre, 2000;Calvi et al, 2003;Zhang et al, 2003;Li and Xie, 2005).…”

Spermatogonial Stem Cells and Animal Transgenesis

“…It was first reported that SSCs can be in vitro cultured for months by Nagano et al (1998). The same group later suggested the addition of GDNF is important to short-term in vitro maintenance of SSCs (Nagano et al, 2003).…”

“…The co-culture of SSCs with monolayers of other cell types as mitotically inactivated murine embryonic fibroblasts (MEF) or STO feeder cells (Nagano et al, 1998) are still discussible, presenting different results so far. MEF is widely used in in vitro culture of embryonic stem cells (Evans & Kaufman, 1981) and has been applied as feeder cells to murine SSCs cultures (Kanatsu-Shinohara et al, 2004a).…”

Effects of extracellular matrices and lectin Dolichos biflorus agglutinin on cell adhesion and self-renewal of bovine gonocytes cultured in vitro

“…However, in domestic animals, little is known about whether gonocytes have stem cell activity during germ cell development. Culture conditions for maintaining germ cells have been established for various species, including the mouse (Nagano et al 1998(Nagano et al , 2003Kubota et al 2004;Kanatsu-Shinohara et al 2005), rat (Hamra et al 2005), hamster (Kanatsu-Shinohara et al 2008a) and rabbit (Kubota et al 2011). However, culture systems are not available for domestic animal species and cell lines, such as embryonic germ (EG) cells in the mouse, have not been established.…”