title: Reconstitution of mouse spermatogonial stem cell niches in culture.
abstract: 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.
authors.affiliation: Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan. mshinoha@virus.kyoto-u.ac.jp
authors.forename: Mito
authors.initials: M
authors.lastname: Kanatsu-Shinohara
authors.forename: Kimiko
authors.initials: K
authors.lastname: Inoue
authors.forename: Seiji
authors.initials: S
authors.lastname: Takashima
authors.forename: Masanori
authors.initials: M
authors.lastname: Takehashi
authors.forename: Narumi
authors.initials: N
authors.lastname: Ogonuki
authors.forename: Hiroko
authors.initials: H
authors.lastname: Morimoto
authors.forename: Takashi
authors.initials: T
authors.lastname: Nagasawa
authors.forename: Atsuo
authors.initials: A
authors.lastname: Ogura
authors.forename: Takashi
authors.initials: T
authors.lastname: Shinohara
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation; D002470:Cell Survival / Q000235:genetics; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000235:genetics / Q000378:metabolism*; D018920:Coculture Techniques; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism*; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism; D012671:Seminiferous Tubules / Q000166:cytology* / Q000254:growth & development; D012708:Sertoli Cells / Q000502:physiology*; D013093:Spermatogonia / Q000502:physiology*; D055153:Stem Cell Niche / Q000502:physiology*; D019076:Transgenes / Q000235:genetics
chemical_list: D054377:Chemokine CXCL12; D051100:Glial Cell Line-Derived Neurotrophic Factor; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; D019718:Receptors, CXCR4