The Impact of Conditioned Medium Derived from Human Adipose-Derived Mesenchymal Stem Cells on the Recovery of Testicular Function in a Rat Model of Azoospermia Induced by Busulfan.
The Impact of Conditioned Medium Derived from Human Adipose-Derived Mesenchymal Stem Cells on the Recovery of Testicular Function in a Rat Model of Azoospermia Induced by Busulfan.
Hassan Hedayatnia,1,*
1. Department of Biology, Qom Branch, Islamic Azad University, Qom, Iran
Introduction: Men are responsible for 20-30% of infertility cases Several factors can contribute to male infertility, including reduced sperm production, abnormal sperm function, obstruction of ejaculatory ducts, and impotence. Meanwhile, non-obstructive azoospermia (NOA), which is characterized as the absence of sperm in ejaculation due to failure of spermatogenesis, is the most severe form of male infertility. Non-obstructive azoospermia is caused by hormonal and genetic disorders, trauma to the testis and its disease, varicocele, etc. Currently, most individuals with non-obstructive azoospermia (NOA) do not have a treatment option that can fully restore spermatogenesis , except for patients with secondary testicular failure. Therefore, the only way for couples to conceive is to receive sperm directly from the testis for intracytoplasmic sperm injection (ICSI). ICSI is the standard treatment for male infertility but, It may increase the risk of genetic abnormalities in the offspings compared to natural pregnancy. Most men with non-obstructive azoospermia (NOA) have no treatment options other than assisted reproductive techniques (ART) to have a biological child. Given current therapies, there is an urgent need to develop alternative and effective therapeutic strategies.Secreted factors by mesenchymal Stem Cells (MSCs) offer promising prospects for cell-based therapy in regenerative medicine. This research is to detect if Conditioned Medium (CM) obtained from Human adipose tissue-derived mesenchymal stem (AD-MSCs) can restore spermatogenesis in busulfan-induced azoospermatic rat model.
Methods: To study the effect of conditioned medium intra-testis injection on the recovery of spermatogenes in NOA rat model ,the adiopose – derived mesenchymal stem cells (Ad-MSCs) were isolated and cultured. Then on the 3rd passage conditioned medium was collected .This study was conducted on 30 male Wistar rats (aged 8-12 weeks). Animals were kept adapted for one week before the beginning of the research in an appropriate environment and had free access to food supplies and water. Rats were divided in 4 groups, as control group ( without treatment or negative control ) , NOA group (busulfan -induced rat or positive control) azoospermia sham (busulfan + Phosphate-buffered saline ) and experimental groups ( busulfan + ADSC-CM).The non-obstructive azoospermia (NOA) model was established using intraperitoneal administration of busulfan in rat by two intraperitoneal injections of busulfan 10 mg/kg, with 21 day intervals. 35 days after the second busulfan injection, the test group was treated with adipose tissue-derived stem cell conditioned medium(500mg/ml).western blot analysis was conducted to assess the levels of DAZL and VASA proteins.
Results: The analysis indicated a significant increase(P ≤ 0:000) in the levels of DAZL and VASA expression within the ADSC-CM group as comparedto the control group, whereas a significant decrease was observed in the NOA and Sham groups.
Conclusion: This research proposes that the use of conditioned medium from human adipose -derived mesenchymal stem cells could potentially aid in the restoration of spermatogenesis in busulfan-induced infertile rats.