Cisplatin Effects on the Human Fetal Testis – Establishing the Sensitive Period for (Pre)Spermatogonial Loss and Relevance for Fertility Preservation in Pre-Pubertal Boys

Abstract

BackgroundExposure to chemotherapy during childhood can impair future fertility. Studies using in vitro culture have shown exposure to platinum-based alkylating-like chemotherapy reduces the germ cell number in the human fetal testicular tissues. We aimed to determine whether effects of exposure to cisplatin on the germ cell sub-populations are dependent on the gestational age of the fetus and what impact this might have on the utility of using human fetal testis cultures to model chemotherapy exposure in childhood testis.MethodsWe utilised an in vitro culture system to culture pieces of human fetal testicular tissues (total n=23 fetuses) from three different gestational age groups (14-16 (early), 17-19 (mid) and 20-22 (late) gestational weeks; GW) of the second trimester. Tissues were exposed to cisplatin or vehicle control for 24 hours, analysing the tissues 72 and 240 hours post-exposure. Number of germ cells and their sub-populations, including gonocytes and (pre)spermatogonia, were quantified.ResultsTotal germ cell number and number of both germ cell sub-populations were unchanged at 72 hours post-exposure to cisplatin in the testicular tissues from fetuses of the early (14-16 GW) and late (20-22 GW) second trimester. In the testicular tissues from fetuses of mid (17-19 GW) second trimester, total germ cell and gonocyte number were significantly reduced, whilst (pre)spermatogonial number was unchanged. At 240 hours post-exposure, the total number of germ cells and that of both sub-populations was significantly reduced in the testicular tissues from fetuses of mid- and late-second trimester, whilst germ cells in early-second trimester tissues were unchanged at this time-point.ConclusionsIn vitro culture of human fetal testicular tissues can be a useful model system to investigate the effects of chemotherapy-exposure on germ cell sub-populations during pre-puberty. Interpretation of the results of such studies in terms of relevance to later (infant and pre-pubertal) developmental stages should take into account the changes in germ cell composition and periods of germ cell sensitivity in the human fetal testis.

Alkylating agents‐induced gonadotoxicity in prepubertal males: Insights on the clinical and preclinical front

“…Another developing aspect of preclinical research is the use of human‐derived tissue. So far, only cisplatin and carboplatin have been tested in such models 89–92 . Possibly due to easier accessibility, research was carried out on fetal tissue after it was found to exhibit a similar pattern and timing of damage to child‐derived tissue following treatment 89 .…”

“…So far, only cisplatin and carboplatin have been tested in such models. [89][90][91][92] Possibly due to easier accessibility, research was carried out on fetal tissue after it was found to exhibit a similar pattern and timing of damage to child-derived tissue following treatment. 89 As expected, this group found some differences when comparing the response of mice and human-derived tissue.…”

“…Subsequently, after slight modifications to the culture media, the authors demonstrated that this method could also support fetal‐derived tissue for 2 weeks 88 . This led another group to rely on this model to investigate the effects of cisplatin and carboplatin treatment on the prepubertal human testis 89–92 . To the best of our knowledge, these are pioneering experiments concerning the use of prepubertal‐aged human tissues in chemotherapy‐induced gonadotoxicity studies and will be further discussed in more detail.…”

“…88 This led another group to rely on this model to investigate the effects of cisplatin and carboplatin treatment on the prepubertal human testis. [89][90][91][92] To the best of our knowledge, these are pioneering experiments concerning the use of prepubertal-aged human tissues in chemotherapy-induced gonadotoxicity studies and will be further discussed in more detail. Despite their novel usage of child-derived tissue, no constraints regarding tissue viability were mentioned.…”

The Effects of White Tea (Camellia Sinensis) and Infliximab Against Cisplatin- Induced Testicular Damage via Oxidative Stress and Apoptosis

“…However, the cisplatin-induced damage in testicular tissue recovered either partially or completely with the administration of white tea and infliximab, both as single agents and in combination. Although there is a multitude of studies showing the occurrence of cisplatin-induced testicular toxicity, the molecular mechanism of cisplatin-related testicular damage has not been completely clarified (Narayana et al, 2009;Aly & Eid, 2020;Matilionyte et al, 2022;Ismail et al, 2023). However, the most widely studied and the most strongly emphasized mechanism is associated with oxidative stress and increased apoptosis (Salem et al, 2012;Azab et al, 2020).…”