Human placental mesenchymal stem cells ameliorate chemotherapy-induced damage in the testis by reducing apoptosis/oxidative stress and promoting autophagy

Abstract

Background The side effects of busulfan on male reproduction are serious, so fertility preservation in children undergoing busulfan treatment is a major worldwide concern. Human placental mesenchymal stem cells (hPMSCs) have advantages such as stable proliferation and lower immunogenicity that make them an ideal material for stimulating tissue repair, especially restoring spermatogenesis. The protective effects of hPMSCs in busulfan-induced Sertoli cells and in busulfan-treated mouse testes have not been determined. Our study aimed to elaborate the protective effect and potential mechanisms of hPMSCs in busulfan-treated testes and Sertoli cells. Methods First, we developed a mouse model of busulfan-induced testicular toxicity in vivo and a mouse Sertoli cell line treated with busulfan in vitro to assess the protective effect and mechanisms of hPMSC treatment on spermatogenesis. Then, the length, width, and weight of the testes were monitored using Vernier calipers. Furthermore, at 1 week and 4 weeks after the transplantation of hPMSCs, histological sections of testes were stained with hematoxylin-eosin, and the seminiferous tubules with fluid-filled cavities were counted. Through ELISA analysis, testosterone levels and MDA, SOD, LDH, and CAT activities, which are associated with ROS, were detected. Markers of ROS, proliferation (Ki67), and apoptosis (Annexin V) were evaluated by FACS. Next, the fluorescence intensity of proliferation markers (BrdU and SCP3), an antioxidant marker (SIRT1), a spermatogenesis marker (PLZF), and autophagy-related genes (P62 and LC3AB) were detected by fluorescence microscopy. The mRNA expression of γ-H2AX, BRCA1, PARP1, PCNA, Ki67, P62, and LC3 was determined by qRT-PCR. Results hPMSCs restored disrupted spermatogenesis, promoted improved semen parameters, and increased testosterone levels, testis size, and autophagy in the testis toxicity mouse model induced by busulfan. hPMSCs suppressed the apoptosis of Sertoli cells and enhanced their rate of proliferation in vitro. Additionally, hPMSCs protected against oxidative stress and decreased oxidative damage in the testis toxicity mouse model induced by busulfan. Furthermore, hPMSCs increased the expression of proliferation genes (PCNA and KI67) and decreased the mRNA levels of apoptotic genes such as γ-H2AX, BRCA1, and PARP1. Conclusions This research showed that hPMSC injection ameliorated busulfan-induced damage in the testis by reducing apoptosis/oxidative stress and promoting autophagy. The present study offers an idea for a new method for clinical treatment of chemotherapy-induced spermatogenesis.

Mesenchymal stromal/stem cells promote intestinal epithelium regeneration after chemotherapy-induced damage

“…The proapoptotic and anti-proliferative effects of busulfan were relatively rapidly reversed by the co-culture with MSCs, suggesting that MSCs release factors, which promote proliferation and inhibit apoptosis. This is in line with previous studies in animal models or in vitro systems using animal cells, where MSCs were shown to rescue radiotherapy-or chemotherapy-mediated damage in organs, such as the testis, colon, or liver [52][53][54][55]. However, the direct anti-apoptotic and pro-proliferative effect of MSCs was only demonstrated in TM4 mouse Sertoli cell line in vitro so far.…”

Regenerative medicine for male infertility: A focus on stem cell niche injury models

“…For some models, it can be assumed that spermatogenesis might be restored due to paracrine stimulation provided by transplanted MSC to maintain the function of injured or dysregulated SSC niche components [ 59 ]. Moreover, the impact of paracrine factors secreted by MSCs on spermatogenesis restoration can be confirmed by the effective restoration of spermatogenesis in animal models of spermatogenesis failure after injection of human MSCs isolated from various tissues or their secretome [ [60] , [61] , [62] ].…”

Cell-Based Therapy Approaches in Treatment of Non-obstructive Azoospermia

“…To date, mesenchymal-like cells isolated from different parts of human placenta including amnion, chorion, and decidua have been used in preclinical and clinical studies to treat various diseases [ 98 , 99 ]. Among these, placenta-derived MSCs (PD-MSCs) have been used for treatment of infertility-related diseases such as premature ovarian failure (POF) [ 100 – 103 ], testicular failure [ 104 ], and male sexual problems such as Peyronie’s disease [ 72 ] and erectile dysfunction (ED) [ 73 ] and have promising results.…”