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
First Page
279
Last Page
297
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.
Recommended Citation
La Salle S,
Sun F,
Handel M.
Isolation and short-term culture of mouse spermatocytes for analysis of meiosis. Methods Mol Biol 2009; 558:279-97.