Short Communication: Isolation and Recognition of Spermatogenic Cell Lineages from Mouse Testis Tissue

• Main Authors: Tahereh Modarresi, Kazem Parivar, Homa Mohseni Kouchesfehani, Marjan Sabbaghian, Hamed Heidari-Vala, Mohammad Reza Sadeghi
• Format: Article
• Language: English
• Published: Royan Institute (ACECR), Tehran 2010-01-01
• Series: Cell Journal
• Subjects: Spermatogenesis; Percoll; Density Gradient Centrifugation; Sedimentation Velocity; Cell Lineage
• Online Access: http://www.celljournal.org/library/upload/article/af_234442615Modaressi.pdf

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