Isolation and enrichment of type A spermatogonia from pre-pubertal buffalo (Bubalus bubalis) testis

T.A. Rafeeqi, G. Kaul

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    7 Citations (Web of Science)

    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. © 2012 Blackwell Verlag GmbH.
    Original languageEnglish
    Pages (from-to)195-203
    JournalAndrologia
    Volume45
    Issue number3
    DOIs
    Publication statusPublished - 2013

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