title: Spermatogonial stem cell enrichment using simple grafting of testis and in vitro cultivation.
abstract: Enrichment of spermatogonial stem cells (SSCs) from the mammalian adult testis faces several limitations owing to their relatively low numbers among many types of advanced germ cells and somatic cells. The aim of the present study was to improve the isolation efficiency of SSCs using a simple tissue grafting method to eliminate the existing advanced germ cells. Sliced testis parenchyma obtained from adult ICR or EGFP-expressing transgenic mice were grafted heterotropically under the dorsal skin of nude mice. The most advanced germ cells disappeared in the grafted tissues after 2-4 weeks. Grafted tissues were dissociated enzymatically and plated in culture dishes. During in vitro culture, significantly more SSCs were obtained from the grafted testes than from non-grafted controls, and the isolated SSCs had proliferative potential and were successfully maintained. Additionally, EGFP-expressing SSCs derived from graft parenchyma were transplanted into bulsufan-treated recipient mice testes. Finally, we obtained EGFP-expressing pups after in vitro fertilization using spermatozoa derived from transplanted SSCs. These results suggest that subcutaneous grafting of testis parenchyma and the subsequent culture methods provide a simple and efficient isolation method to enrich for SSCs in adult testis without specific cell sorting methods and may be useful tools for clinical applications.
authors: Fertility Center of CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul, 135-913, Korea.
authors: Jung Jin
authors: JJ
authors: Lim
authors: Department of Biomedical Science, College of Life Science, CHA University, Seoul, 135-081, Korea.
authors: Dong Won
authors: DW
authors: Seol
authors: Fertility Center of CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul, 135-913, Korea.
authors: Kyung Hee
authors: KH
authors: Choi
authors: Fertility Center of CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul, 135-913, Korea.
authors: Dong Hyuk
authors: DH
authors: Shin
authors: Fertility Center of CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul, 135-913, Korea.
authors: Hyung Joon
authors: HJ
authors: Kim
authors: Department of Urology, CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul, 135-913, Korea.
authors: Seung-Hun
authors: SH
authors: Song
authors: 1] Fertility Center of CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul, 135-913, Korea [2] Department of Biomedical Science, College of Life Science, CHA University, Seoul, 135-081, Korea.
authors: Dong Ryul
authors: DR
authors: Lee
mesh_terms: D053687:Adult Stem Cells / Q000502:physiology*; D000818:Animals; D018929:Cell Culture Techniques; D002469:Cell Separation; D002478:Cells, Cultured; D008297:Male; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D013093:Spermatogonia / Q000502:physiology*; D013737:Testis / Q000166:cytology / Q000637:transplantation*