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Spermatogenesis

Spermatogenesis

Spermatogenesis is the process by which haploid sperm cells are produced in the male testes. It starts with stem cells located close to the outer rim of seminiferous tubules. These spermatogonial stem cells divide asymmetrically to give rise to additional stem cells (meaning that these structures “self-renew”), as well as sperm progenitors, called spermatocytes. Importantly, this method of asymmetric mitotic division maintains a population of spermatogonial stem cells in the male reproductive...

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Related Experiment Video

Updated: May 30, 2026

Functional Assessment of Kinesin-7 CENP-E in Spermatocytes Using In Vivo Inhibition, Immunofluorescence and Flow Cytometry

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Dynamic changes in EPCAM expression during spermatogonial stem cell differentiation in the mouse testis.

Mito Kanatsu-Shinohara¹, Seiji Takashima, Kei Ishii

¹Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto, Japan.

|August 23, 2011

Summary

Spermatogonial stem cells (SSCs) are best purified from CD9(+)EPCAM(low/-) populations. Epithelial cell adhesion molecule (EPCAM) is crucial for progenitor cell amplification in mouse spermatogenesis, aiding SSC purification strategies.

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Transillumination-Assisted Dissection of Specific Stages of the Mouse Seminiferous Epithelial Cycle for Downstream Immunostaining Analyses

Published on: October 7, 2020

Area of Science:

Reproductive Biology
Stem Cell Biology
Developmental Biology

Background:

Spermatogonial stem cells (SSCs) possess self-renewal capabilities but are morphologically indistinguishable from committed spermatogonia.
Identifying reliable markers for SSC purification is crucial for research and therapeutic applications.
CD9 and epithelial cell adhesion molecule (EPCAM) are known markers also found on embryonic stem cells.

Purpose of the Study:

To analyze the expression patterns of CD9 and EPCAM in relation to SSC activity.
To establish a method for distinguishing SSCs from committed spermatogonia.
To develop an improved purification strategy for SSCs from testicular cells.

Main Methods:

Magnetic cell sorting (MACS) and fluorescence-activated cell sorting (FACS) were employed to isolate and analyze germ cell populations.
Expression levels of CD9 and EPCAM were correlated with SSC clonogenic potential and activity.
Short hairpin RNA (shRNA) was used to suppress EPCAM expression in germline stem (GS) cell cultures.

Main Results:

EPCAM-selected mouse germ cells exhibited limited in vitro clonogenic potential.
The CD9(+)EPCAM(low/-) population showed a significantly higher frequency of SSCs compared to the CD9(+)EPCAM(+) population.
EPCAM suppression compromised GS cell proliferation and increased SSC concentration, while overexpression did not significantly affect SSC activity.

Conclusions:

SSCs are most concentrated within the CD9(+)EPCAM(low/-) cell population.
EPCAM plays a significant role in the amplification of progenitor cells within the mouse spermatogenic system.
Distinguishing progenitor spermatogonia from SSCs is key to refining SSC purification methods.