title: Isolation and enrichment of type A spermatogonia from pre-pubertal buffalo ( Bubalus bubalis) testis.
abstract: The aim of this study was to isolate and subsequently enrich type A spermatogonia from pre-pubertal buffalo testis. Two-step enzymatic digestion was used to isolate spermatogonia from 10 to 14 months pre-pubertal buffalo calves, resulting in maximal release of spermatogonia from the seminiferous tubules. After enzymatic digestion, the type A spermatogonia were subsequently enriched by differential plating and Percoll gradient centrifugation. The identity of type A spermatogonia was determined by light microscopy and further characterised by Dolichos biflorus agglutinin, a specific marker for bovine type A spermatogonia by fluorescence-activated cell sorting analysis. After enzymatic isolation, the cell suspension contained about 27% of type A spermatogonia, which was enriched up to 71% with >70% cell viability. Further flow cytometric analysis showed the presence of THY1+ cells (cells expressing thymocyte differentiation antigen 1), suggesting that THY1 is a conserved marker of the undifferentiated spermatogonial cells in buffalo. The isolation of the enriched type A spermatogonia from buffalo testis opens ways to study the further biochemical characteristics of this important class of germ cells in this species.
authors: Department of Biochemistry, National Dairy Research Institute, I.C.A.R., Karnal-132001, Haryana, India.
authors: T
authors: T
authors: Rafeeqi
authors: G
authors: G
authors: Kaul
mesh_terms: D000818:Animals; D002020:Buffaloes / Q000254:growth & development / Q000502:physiology*; D002417:Cattle; D002470:Cell Survival; D005434:Flow Cytometry; D008297:Male; D037121:Plant Lectins; D055101:Semen Analysis / Q000662:veterinary*; D012741:Sexual Maturation / Q000502:physiology*; D013091:Spermatogenesis / Q000502:physiology; D013093:Spermatogonia / Q000166:cytology*; D013737:Testis / Q000166:cytology* / Q000254:growth & development*
chemical_list: D037121:Plant Lectins; C036554:dolichos biflorus agglutinin