title: Purification and Analysis of Male Germ Cells from Adult Mouse Testis.
abstract: Isolation of pools of spermatogenic cells at specific developmental stages is essential for the investigations of molecular events controlling critical transitions during spermatogenesis. Large-scale cell purification techniques allow for combined proteomics, genomics, and transcriptomics studies. Herein, we describe a procedure for the purification of meiotic and post-meiotic male germ cells from adult mouse testes. We also describe how the fractionated cell populations could be used for further studies. In our laboratory, these protocols are routinely used to specifically investigate the molecular basis of histone acetylation/acylation-driven epigenetic programming.
authors.affiliation: CNRS UMR 5209, INSERM U1209, CR UGA, Institute for Advanced Biosciences, La Tronche, 38700, France. thierry.buchou@univ-grenoble-alpes.fr.
authors.forename: Thierry
authors.initials: T
authors.lastname: Buchou
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China.
authors.forename: Minjia
authors.initials: M
authors.lastname: Tan
authors.affiliation: CNRS UMR 5209, INSERM U1209, CR UGA, Institute for Advanced Biosciences, La Tronche, 38700, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Barral
authors.affiliation: CNRS UMR 5209, INSERM U1209, CR UGA, Institute for Advanced Biosciences, La Tronche, 38700, France.
authors.forename: Anne-Laure
authors.initials: AL
authors.lastname: Vitte
authors.affiliation: CNRS UMR 5209, INSERM U1209, CR UGA, Institute for Advanced Biosciences, La Tronche, 38700, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Rousseaux
authors.affiliation: Laboratory of Stem Cells, Development and Cancer, Department of Cell Biology and Histology, University of the Basque Country (UPV/EHU), Vizcaya, Spain.|Analytical and High Resolution Biomedical Microscopy Core Facility, University of the Basque Country (UPV/EHU), Vizcaya, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Arechaga
authors.affiliation: CNRS UMR 5209, INSERM U1209, CR UGA, Institute for Advanced Biosciences, La Tronche, 38700, France.
authors.forename: Saadi
authors.initials: S
authors.lastname: Khochbin
mesh_terms: D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D047369:Chromatin Immunoprecipitation; D044127:Epigenesis, Genetic; D005466:Fluorocarbons / Q000737:chemistry; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006655:Histone Deacetylases / Q000235:genetics* / Q000378:metabolism; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D006877:Hydroxamic Acids / Q000494:pharmacology; D008297:Male; D008540:Meiosis; D051379:Mice; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013087:Spermatids / Q000166:cytology* / Q000378:metabolism; D013090:Spermatocytes / Q000166:cytology* / Q000378:metabolism; D013091:Spermatogenesis / Q000235:genetics; D013737:Testis / Q000166:cytology / Q000254:growth & development / Q000378:metabolism
chemical_list: D005466:Fluorocarbons; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids; C429307:fluorocarbon 77; D012710:Serum Albumin, Bovine; C012589:trichostatin A; D006655:Histone Deacetylases
keywords: Haploid post-meiotic spermatid; Male germ cell; Mouse testis