In Vitro Effect of Human Chorionic Gonadotropin Treatment on Gubernaculum and Testis Cell Stimulation and Vascularization: The Role of Therapy in Testicular Disease

Background: Cryptorchidism is a disease with a significant incidence among newborns and has two main long-term complications, such as infertility and testicular cancer. Surgery represents the treatment of choice, preferably performed within 2-3 years of the patient's life. In order to improve the postoperative results, especially regarding the functional recovery of the gonad, human chorionic gonadotropin (hCG) treatment is proposed as postoperative adjuvant therapy, due its clinical efficacy previously demonstrated in different studies. The aim of this study is to understand the molecular rationale of the treatment for each patient by evaluating clinical aspects together with molecular analysis performed on patient’s derived tissue.

Methods: To perform this study, we took advantage of the availability of primary testicular and gubernaculum tissues derived from pediatric patients affected by cryptorchidism combined with expertise in cell culture and molecular biology. Indeed, testicular and gubernaculum cells derived from a small portion of tissue (abut 10-20 mm3) obtained during orchidopexy have been cultured and amplified in vitro by following a protocol optimized by our laboratory. When the cells achieved exponential growth in vitro, molecular studies and analysis of cellular response to hCG hormone treatment were performed for each individual patient. The time and doses of hCG in vitro treatment have been considered as similar as possible to that provided in clinical practice.

Results: In this project, we demonstrate for the first time in pediatric patients that a specific disruption of gubernaculum and testis parenchyma biopsies allows to obtain viable primary cells that possess the ability to grow in culture (Figure 1). The results obtained on in vitro cultured primary cells show that they express at mRNA and protein levels the hCG-specific receptor (LHR) and that the use of hCG treatment significantly stimulates the in vitro proliferation of gubernaculum and Leydig cells compared to untreated cells from the same patient (Figure 2).

Conclusion: The obtained results showing that hCG increases the proliferative capacity of gubernaculum and testis cells cultured in vitro offer the possibility to compare the in vitro response with the increase in testicular volume in the patient following treatment. The potential of this study is of great impact as it opens the way to the evaluation of cellular response to different dosages of hCG hormone and, therefore, to design an effective personalized therapy for each individual patient.