title: Spermatogonia-specific proteins expressed in prepubertal buffalo (Bubalus bubalis) testis and their utilization for isolation and in vitro cultivation of spermatogonia.
abstract: Buffalo is an economically important livestock species in Asia. Little is known about male germ line technology owing to lack of sufficient understanding regarding expression of germ- and somatic-cell specific-proteins in the testis. In this study, we identified UCHL-1 (PGP 9.5) and lectin- Dolichos biflorus agglutinin (DBA) as specific markers for spermatogonia in buffalo testis. Expression of germ-cell and pluripotency-specific proteins such as DDX4 (VASA) and POU5F1 (OCT3/4) were also present in spermatogonia. Interestingly, the expression of somatic cell-specific proteins such as VIMENTIN and GATA4 were also detected in germ cells. Using two-step enzymatic digestion followed by differential plating and Percoll density-gradient centrifugation, an approximately 55% spermatogonia-enriched cell population could be obtained from the prepubertal buffalo testis. Isolated spermatogonia could survive and proliferate in vitro in DMEM/F12 medium containing 10% fetal bovine serum in the absence of any specific growth factors for a week. Cultured spermatogonia showed DBA affinity and expressed DDX4 and POU5F1. These results may help to establish a long-term culture system for buffalo spermatogonia.
authors: Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India. sandeep@ccmb.res.in
authors: Sandeep
authors: S
authors: Goel
authors: Niranjan
authors: N
authors: Reddy
authors: Suman
authors: S
authors: Mandal
authors: Mayako
authors: M
authors: Fujihara
authors: Sung-Min
authors: SM
authors: Kim
authors: Hiroshi
authors: H
authors: Imai
mesh_terms: D000818:Animals; D002020:Buffaloes / Q000502:physiology*; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D005786:Gene Expression Regulation / Q000502:physiology*; D008297:Male; D012741:Sexual Maturation / Q000502:physiology*; D013093:Spermatogonia / Q000166:cytology / Q000502:physiology*; D013737:Testis / Q000378:metabolism*