Title

Isolation and short-term culture of mouse spermatocytes for analysis of meiosis.

Document Type

Article

Publication Date

2009

Keywords

Animals, Cell Culture Techniques, Cell Separation, Cells, Cultured, Cytogenetic Analysis, Male, Meiosis, Mice, Models, Biological, Spermatocytes, Time Factors

Publisher

Humana Press

JAX Location

Reprint Collection

JAX Source

Methods Mol Biol 2009; 558:279-97.

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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