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Step-specific Sorting of Mouse Spermatids by Flow Cytometry
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Practical selection methods for rat and mouse round spermatids without DNA staining by flow cytometric cell sorting.

Tomonari Hayama1, Tomoyuki Yamaguchi1, Megumi Kato-Itoh1

  • 1Division of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo, Japan.

Molecular Reproduction and Development
|April 1, 2016
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Summary

Researchers developed a simple method using flow cytometry to isolate round spermatids for round spermatid injection (ROSI). This technique enables fertility treatments for males with severe spermatogenesis disorders, leading to healthy offspring.

Keywords:
ROSIflow cytometrymouserat

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Area of Science:

  • Reproductive Biology
  • Cell Biology
  • Genetics

Background:

  • Severe spermatogenesis disorders can cause male infertility.
  • Round spermatid injection (ROSI) offers a potential treatment, but isolating round spermatids is challenging.
  • Efficient isolation of round spermatids is crucial for clinical application.

Purpose of the Study:

  • To develop an efficient and simple method for isolating rodent haploid round spermatids.
  • To assess the efficacy of flow cytometry for round spermatid isolation and subsequent ROSI.

Main Methods:

  • Utilized flow cytometric cell sorting to isolate round spermatids.
  • Separation was based on DNA content (Hoechst 33342 or Dye Cycle Violet) or cell characteristics (forward/side scatter).
  • Performed ROSI using flow cytometry-selected round spermatids.

Main Results:

  • Successfully isolated rodent haploid round spermatids using flow cytometry.
  • Achieved healthy offspring following ROSI with flow cytometry-selected round spermatids.
  • Demonstrated the effectiveness of the non-invasive isolation method.

Conclusions:

  • Flow cytometry provides an efficient and simple method for isolating round spermatids.
  • This technique is a viable option for human male infertility treatment and animal breeding.
  • The method facilitates successful ROSI, leading to healthy offspring.