Abstract
Understanding meiosis is facilitated by in vitro experimental approaches, but this has not been easily applicable to mammalian meiocytes. Available methods for in vitro analysis of mammalian oocytes are generally limited to experimental analysis of the late prophase period. Short-term cultures of male germ cells have been useful for analysis of earlier meiotic prophase pathways, as well as onset of the meiotic division phase, but no studies have achieved reliable spermatogenesis in vitro. Here we describe a method for preparing highly enriched pachytene spermatocytes from mouse testicular cell suspensions using cell-size fractionation by sedimentation through a bovine serum albumin gradient at unit gravity. We also provide a procedure for short-term culture of spermatocytes and the pharmacological induction of the prophase-to-division phase transition.
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Acknowledgments
Work described here is supported by NIH grants to M.A.H. (HD33816 and HD48998) and a Cancer Center Core Grant to The Jackson Laboratory (CA34196). S.L. is funded in part by a fellowship from les Fonds de la Recherche en Santé du Québec (FRSQ).
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La Salle, S., Sun, F., Handel, M.A. (2009). Isolation and Short-Term Culture of Mouse Spermatocytes for Analysis of Meiosis. In: Keeney, S. (eds) Meiosis. Methods in Molecular Biology, vol 558. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-103-5_17
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DOI: https://doi.org/10.1007/978-1-60761-103-5_17
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