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Related Experiment Video

Updated: Aug 27, 2025

Isolation of Sertoli Cells and Peritubular Cells from Rat Testes
11:11

Isolation of Sertoli Cells and Peritubular Cells from Rat Testes

Published on: February 8, 2016

22.7K

Two complementary approaches for efficient isolation of Sertoli cells for transcriptomic analysis.

Jana Petrusová1, Jasper Manning1, Jan Kubovčiak2

  • 1Laboratory of Immunobiology, Institute of Molecular Genetics of the Czech Academy of Sciences, Prague, Czechia.

Frontiers in Cell and Developmental Biology
|September 26, 2022
PubMed
Summary

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Researchers developed a rapid, 3-hour method using fluorescence-activated cell sorting (FACS) to isolate pure Sertoli cells (SCs) for studying male fertility and spermatogenesis.

Area of Science:

  • Reproductive Biology
  • Cell Biology
  • Spermatogenesis Research

Background:

  • Sertoli cells (SCs) are crucial somatic cells supporting germ cell development and spermatogenesis within seminiferous tubules.
  • SC maturation is vital for male fertility, and disruptions can lead to reproductive deficiencies.
  • A precise and rapid method for isolating pure, viable SCs has been lacking, hindering research.

Purpose of the Study:

  • To develop and validate improved methods for rapid, high-purity isolation of viable Sertoli cells (SCs) from testicular tissue.
  • To enable accurate analysis of the SC transcriptome for understanding spermatogenesis and male infertility.

Main Methods:

  • Developed two fluorescence-activated cell sorting (FACS) protocols for SC isolation.
  • Protocol 1: Sorting based on co-expression of FSH receptor (FSHr) and Occludin-1.
Keywords:
FSHrOccludinSertoli cellflow cytometryspermatogenesis

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  • Protocol 2: Sorting based on FSHr antibody co-staining and Hoechst 33342 DNA content discrimination.

    • Main Results:

    • The entire isolation procedure takes less than 3 hours, allowing for timely transcriptional analysis.
    • The developed FACS methods yield SC transcriptomes comparable to those from fluorescent protein-tagged SCs.
    • Sorting by 2C DNA content effectively minimized germ cell-associated transcripts, offering a more accurate SC transcriptome.

    Conclusions:

    • Novel FACS-based approaches provide a reliable and rapid means for isolating pure, viable Sertoli cells.
    • These methods will facilitate research into SC function, spermatogenesis, and male infertility disorders.
    • Sorting SCs by DNA content is recommended for precise transcriptome analysis.