Isolation And Recognition Of Spermatogenic Cell Lineages From Mouse Testis Tissue

Authors

1 Department of Biology, Tarbiat Moallem University, Tehran, Iran

2 Department of Andrology, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

3 Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran

4 Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran

Abstract

The main objective of this study was to compare the quality and effectiveness of the sedimentation velocity at unit gravity (SVUG) method with the percoll density gradient centrifugation method in the separation of spermatogenic cells in order to introduce a simple and cost-effective method. In this study, the testes from male mice (NMRI strain) were obtained and transferred to Ham's F10 medium. With the use of the mechanical and enzymatic digestion method, spermatogenic cells were isolated. Spermatogenic cell separation was undertaken with both the percoll gradient centrifugation and sedimentation velocity at unit gravity (SVUG) methods based on human serum albumin (HSA) gradients. Subsequently the May-Grunwald-Giemsa (MGG) staining method was used to identify isolated cells. The combined method possibly could isolate a purer fraction of spermatogenic cells. This method (SVUG and Percoll Gradients Centrifugation combination) has the potential to be used as a simple and effective method for the isolation of spermatogenic cells for diagnostic or research purposes.

Keywords

Cell Journal (Yakhteh)
Volume 12, Issue 3
September 2010
Pages 429-432

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