Related Experiment Video
Updated: Nov 22, 2025
A Seminiferous Tubule Squash Technique for the Cytological Analysis of Spermatogenesis Using the Mouse Model
Published on: February 6, 2018
Chemotherapy induced damage to spermatogonial stem cells in prepubertal mouse in vitro impairs long-term
Federica Lopes1,2, Prathima Tholeti1,3, Satish K Adiga3
1Biomedical Sciences, University of Edinburgh, Edinburgh, UK.
Abstract:
Chemotherapy can affect testis development of young boys with cancer, reducing the chances of fatherhood in adulthood. Studies using experimental models are needed to determine the damage caused by individual chemotherapy drugs in order to predict the risk of infertility and direct patients towards appropriate fertility preservation options. Here, we investigated the individual role of two drugs, cisplatin and doxorubicin, using an in vitro culture model of prepubertal (postnatal day 5) mouse testis that supports induction and maintenance of full spermatogenesis. Twenty-four hour exposure with either drug at clinically-relevant doses (0.25, 0.5 or 0.75 microg/mL for cisplatin, or 0.01, 0.03 or 0.05 microg/mL for doxorubicin), induced an acute significant loss of spermatogonial stem cells (SSCs; PLZF+), proliferating SSCs (PLZF+BrdU+), total germ cells (MVH+), and spermatocytes (SCP3+) one week after chemotherapy exposure. By the time of the first (Week 4) and second (Week 8) waves of spermatogenesis, there was no longer any effect on SSC or proliferating SSC numbers in drug-exposed testis compared to untreated tissue: however, the populations of total germ cells and spermatocytes were still lower in the higher-dose cisplatin treated groups, along with a reduced frequency of round and elongated spermatids in both cisplatin- and doxorubicin-treated testis fragments. Overall, this study details a direct impairment of germ cell development following acute chemotherapy-induced damage during the prepubertal phase, most likely due to an effect on SSCs, using an in vitro culture system that successfully recapitulates key events of mouse spermatogenesis.
Insights
Chemotherapy drugs like cisplatin and doxorubicin can harm young boys' fertility by damaging sperm development. This study used an in vitro model to show how these drugs affect spermatogonial stem cells and germ cell populations.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Toxicology
Background:
- Chemotherapy poses a risk to testicular function and future fertility in young male cancer survivors.
- Understanding the specific impact of individual chemotherapy agents on prepubertal testis development is crucial for fertility preservation strategies.
- Experimental models are needed to assess drug-induced testicular damage and predict infertility risks.
Purpose of the Study:
- To investigate the individual effects of cisplatin and doxorubicin on prepubertal mouse testis development.
- To assess the impact of these chemotherapy drugs on spermatogonial stem cells (SSCs) and subsequent germ cell populations in vitro.
- To evaluate the long-term consequences on spermatogenesis following acute chemotherapy exposure.
Main Methods:
- Utilized an in vitro culture model of postnatal day 5 mouse testis.
- Exposed testis fragments to clinically relevant doses of cisplatin or doxorubicin for 24 hours.
- Quantified spermatogonial stem cells (SSCs), proliferating SSCs, total germ cells, and spermatocytes at various time points post-exposure.
Main Results:
- Acute exposure to cisplatin and doxorubicin caused significant loss of SSCs, proliferating SSCs, total germ cells, and spermatocytes one week post-treatment.
- While SSC numbers recovered, higher doses of cisplatin and both drugs reduced total germ cells and spermatocytes by weeks 4 and 8.
- Reduced frequencies of round and elongated spermatids were observed in both cisplatin- and doxorubicin-treated testis fragments.
Conclusions:
- Acute chemotherapy exposure during the prepubertal phase directly impairs germ cell development, primarily affecting spermatogonial stem cells.
- The in vitro model successfully recapitulates key aspects of chemotherapy-induced testicular damage and spermatogenesis.
- Findings highlight the potential for long-term fertility impairment due to chemotherapy in young males.
Related Concept Videos
01:41Spermatogenesis
01:22Spermatogenesis
The process of spermatogenesis can be divided into mitosis, meiosis, and spermiogenesis. During mitosis, the spermatogonia or stem cells divide to produce two identical daughter cells, type A and B spermatogonia. Type-A...