Immunolocalization of Steroidogenic Enzymes (3β-hydroxysteroid dehydrogenase, 17β-hydroxysteroid dehydrogenase, and P450scc) in Rats with Testicular Dysfunction Treated with Mesenchymal Stem Cells-conditioned Medium

About 60-80 million couples in the world are suffering from infertility disease. Infertility is a major problem in
patients coping with chemotherapy. The chemotherapy process can degenerate non-target organs, especially in
testes. Infertility in male or testicular dysfunction is caused by the failure of proliferation and differentiation of the spermatogenic cells. Many studies reported that mesenchymal stem cells-conditioned medium promoted
regenerative processes. The present study aimed to investigate the effect of mesenchymal stem cells-conditioned
medium on the cisplatin-induced testicular dysfunction by examining the immunolocalization of steroidogenic
enzymes, such as 3β-hydroxysteroid dehydrogenase, 17β-hydroxysteroid dehydrogenase, and P450scc which are
considered as markers of steroid production. All experimental animals were divided into three groups, namely the control group, mesenchymal stem cells-conditioned medium treated group with an injection dose of 0.2 ml/kg body
weight (BW, P1), and mesenchymal stem cells-conditioned medium treated group with an injection dose of 0.5
ml/kg BW (P2). Cisplatin was injected into both treated groups to induce testicular dysfunction. The testicular
tissues were processed by the paraffin method, then cut to a thickness of 5 µm, followed by immunohistochemical
staining. The HSD3B1 immunoreactivities were found only in Leydig cells, and the intensity increased every week
after the injection of mesenchymal stem cells-conditioned medium. The variety of weeks and groups was
significantly different in the number of immunoreactive cells of HSD3B1. The results indicated a significant
difference between one week after the first injection and the one week after the third and fourth injection. The
findings showed a significant difference between the treated group with an injection dose of 0.2 ml/kg BW and the
control group. The number of immunoreactive cells of HSD3B1 with an injection dose of 0.5 ml/kg BW was greater
compared to the group that received an injection dose of 0.2 ml/kg BW. The intensity of HSD3B1 and HSD17B1
increased every week. The p450scc immunoreactive cells were only found in Leydig cells. The intensity of positive
cells of p450scc in the treated group with an injection dose of 0.5 ml/kg BW was more intense, compared to the
treated group with an injection dose of 0.2 ml/kg BW. The results of the current study showed that the injection of
mesenchymal stem cells-conditioned medium can improve the regeneration of spermatogenic cells, and recover
spermatogenesis proved by positive cells of HSD3B1, HSD17B1, and p450scc as markers of steroid production