Isolating mitotic and meiotic germ cells from male mice by developmental synchronization, staging, and sorting

Developmental Biology
Katherine A RomerDavid C Page

Abstract

Isolating discrete populations of germ cells from the mouse testis is challenging, because the adult testis contains germ cells at every step of spermatogenesis, in addition to somatic cells. We present a novel method for isolating precise, high-purity populations of male germ cells. We first synchronize germ cell development in vivo by manipulating retinoic acid metabolism, and perform histological staging to verify synchronization. We use fluorescence-activated cell sorting to separate the synchronized differentiating germ cells from contaminating somatic cells and undifferentiated spermatogonia. We achieve ~90% purity at each step of development from undifferentiated spermatogonia through late meiotic prophase. Utilizing this "3 S" method (synchronize, stage, and sort), we can separate germ cell types that were previously challenging or impossible to distinguish, with sufficient yield for epigenetic and biochemical studies. 3 S expands the toolkit of germ cell sorting methods, and should facilitate detailed characterization of molecular and biochemical changes that occur during the mitotic and meiotic phases of spermatogenesis.

Citations

Nov 2, 2019·Andrology·Rachel GewissMichael D Griswold
Dec 2, 2020·Molecular & Cellular Proteomics : MCP·Kailun FangCharlie Degui Chen
Jan 29, 2021·International Journal of Molecular Sciences·Rosana Rodríguez-Casuriaga, Adriana Geisinger
Feb 23, 2021·EMBO Reports·Enrique AlfaroRocío Gómez
Mar 23, 2021·Frontiers in Cell and Developmental Biology·Adriana GeisingerRicardo Benavente
Nov 14, 2021·Nature Communications·Debashish U MenonTerry Magnuson
Feb 8, 2022·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Mark E GillAntoine H F M Peters

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