Intra-testicular injection of autologous extracellular vesicle from adipose tissue mesenchymal stem cell (EV-ADMSC) in non-obstructive azoospermia Mice

Background: Azoospermia affects about 1% of the male population and may be seen in around 20% of male infertility conditions. In testicular azoospermia the testes are abnormal, atrophic, or absent, and sperm production severely disturbed to absent. FSH levels tend to be elevated (hypergonadotropic) as the feedback loop is interrupted. The condition is seen in 49-93% of men with azoospermia. Non-obstructive azoospermia (NOA) is generally considered a non-medically manageable cause of male infertility. A number of in vitro studies confirmed that mesenchymal stem cells (MSCs) could differentiate into germ cells. MSCs injection reconstructed testicular germinal epithelium in infertile male animal models. In addition, some studies reported that MSCs could differentiate into sperm and regenerate spermatogenesis. There’s is a limited number of published clinical trials in this regard which showed the promising results of this approach in treating male infertility.Objective: We hypothesized that Extracellular Vesicle from Adipose Tissue Mesenchymal Stem Cell (EV-ADMSC) are useful in treating NOA.Materials and Methods: Adult male mice were randomly divided into three groups including the intact control group, azoospermia-induced group, and Conditioned Media (CM) of Adipose Tissue Mesenchymal Stem Cells (AT-MSCs) therapy. Except intact control the other groups were intraperitoneally received two doses of 10 mg/kg of busulfan with 21 days’ interval in order to induce azoospermia. AT-MSCs were isolated from subcutaneous fat tissue of donor mice. The CM of AT-MSCs were injected into the efferent tubules of testes of treatment group 35 days after the last busulfan injection. The cell therapy groups were sampled 60 days after cell therapy and the azoospermia group and azoospermia and control groups were sampled in the same date. The histomorphometric indices of testes were evaluated.Results: Histopathologic evaluation of testes showed that most of the seminiferous tubules of CM therapy group had normal morphology and showed spermatogenesis. Spermatogenesis was not observed in the azoospermia group.Conclusion: CM of AT-MSCs of mice recovered spermatogenesis in the seminiferous tubules of busulfan-induced azoospermic testes. Therefore, CM of AT-MSCs can be suggested as a candidate in CM therapy of azoospermia.

Keywords:

Non-obstructive azoospermia (NOA) , Extracellular vesicle , Mesenchymal Stem Cells , Spermatogenesis