Reconstitution of Mouse Spermatogonial Stem Cell Niches in Culture

• Published in: Cell stem cell 2012-10, Vol.11 (4), p.567-578
• Main Authors: Kanatsu-Shinohara, Mito, Inoue, Kimiko, Takashima, Seiji, Takehashi, Masanori, Ogonuki, Narumi, Morimoto, Hiroko, Nagasawa, Takashi, Ogura, Atsuo, Shinohara, Takashi
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Cell differentiation, maturation, development, hematopoiesis; Cell Movement - genetics; Cell physiology; Cell Proliferation; Cell Survival - genetics; Cells, Cultured; Chemokine CXCL12 - genetics; Chemokine CXCL12 - metabolism; Coculture Techniques; Fundamental and applied biological sciences. Psychology; Glial Cell Line-Derived Neurotrophic Factor - metabolism; Glial Cell Line-Derived Neurotrophic Factor Receptors - genetics; Glial Cell Line-Derived Neurotrophic Factor Receptors - metabolism; Male; Mice; Mice, Inbred C57BL; Molecular and cellular biology; Receptors, CXCR4 - genetics; Receptors, CXCR4 - metabolism; Seminiferous Tubules - cytology; Seminiferous Tubules - growth & development; Sertoli Cells - physiology; Spermatogonia - physiology; Stem Cell Niche - physiology; Transgenes - genetics
• ISSN: 1934-5909; 1875-9777
• DOI: 10.1016/j.stem.2012.06.011

Spermatogonial stem cells (SSCs) reside in specific niches within seminiferous tubules. These niches are thought to secrete chemotactic factors for SSCs, because SSCs migrate to them upon transplantation. However, the identity of these chemotactic molecules remains unknown. Here, we established a testis feeder cell culture system and used it to identify SSC chemotactic factors. When seeded on testis cells from infertile mice, SSCs migrated beneath the Sertoli cells and formed colonies with a cobblestone appearance that were very similar to those produced by hematopoietic stem cells. Cultured cells maintained SSC activity and fertility for at least 5 months. Cobblestone colony formation depended on GDNF and CXCL12, and dominant-negative GDNF receptor transfection or CXCL12 receptor deficiency reduced SSC colonization. Moreover, GDNF upregulated CXCL12 receptor expression, and CXCL12 transfection in Sertoli cells increased homing efficiency. Overall, our findings identify GDNF and CXCL12 as SSC chemotactic factors in vitro and in vivo. [Display omitted] ► Spermatogonial stem cells (SSCs) retain activity in primary testis cell coculture ► SSCs migrate to the in vitro niche and form cobblestone colonies ► GDNF and CXCL12 enhance SSC migration and cobblestone colony formation ► SSC niche colonization in vivo requires CXCL12 and its receptor CXCR4 Coculture of spermatogonial stem cells with testis tissue reconstitutes the niche and highlights a key role for GDNF and CXCL12 in stem cell homing.