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A Standardized Approach for Multispecies Purification of Mammalian Male Germ Cells by Mechanical Tissue Dissociation and Flow Cytometry
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1Department of Genetics,Washington University School of Medicine, 2Graduate Program in Areas of Basic and Applied Biology (GABBA), Abel Salazar Institute of Biomedical Sciences,University of Porto, 3Instituto de Investigação e Inovação em Saúde,University of Porto, 4IPATIMUP – Instituto de Patologia e Imunologia Molecular da Universidade do Porto, 5Department of Pathology & Immunology,Washington University School of Medicine
Chapters
- 00:05Title
- 01:10Testicular Tissue Single Cell Suspension Generation
- 02:26Cell Staining and Fluorescence Activiated Cell Sorting (FACS)
- 05:33Results: Representative Testicular Cell Subset Analyses
- 07:17Conclusion
This work describes the standardization of a method to obtain purified germ cell populations from testicular tissue of different mammalian species. It is a straightforward protocol that combines mechanical testis dissociation, staining with Hoechst-33342 and propidium iodide, and FACS sorting, with wide applications in comparative studies of male reproductive biology.
