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Developmental Biology

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JoVE Journal Developmental Biology
A Standardized Approach for Multispecies Purification of Mammalian Male Germ Cells by Mechanical Tissue Dissociation and Flow Cytometry
 

A Standardized Approach for Multispecies Purification of Mammalian Male Germ Cells by Mechanical Tissue Dissociation and Flow Cytometry

Article DOI: 10.3791/55913-v • 08:46 min • July 12th, 2017
July 12th, 2017 •
*1,2,3,4, *1, 1, 1, 3,4, 1
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
* These authors contributed equally

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résumé
Automatic Translation

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.

Tags

Standardized Approach Multispecies Purification Mammalian Male Germ Cells Mechanical Tissue Dissociation Flow Cytometry Molecular Study Spermatogenesis High-throughput Analysis Germ Cell Types Reagents Mammalian Species Gene Expression Regulation Cell Sorter Gates Decapsulation Tissue Pieces Tissue Disaggregation Cartridge DMEM Medium Tissue Disaggregation System Cell Suspension
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