Abstract
To clarify the effect of busulfan on the depletion of spermatogonial stem cells (SSCs) from shal rams testis, in the first experiment, lambs were treated by intraperitoneal injection with 4 mg/kg busulfan. In the second experiment, different concentrations of busulfan (1, 2 and 4 mg/kg) were injected directly into both sides of the left testis. The testes of 8 lambs were collected by standard castration procedure for histological analysis five weeks after the treatments and the left testis of remaining lambs were collected after eight weeks and a two-time enzymatic digestion process was used to isolate SSCs. The results showed that all rams that had received intraperitoneal injections of busulfan died. But by testicular injecting of same dose of the drug, 40% of the animals died. The testicular injection of rams with 1, 2 and 4 mg/kg of busulfan resulted in a dose dependent decrease in testis size and also spermatocytes population after 5 weeks of treatments. From the results of colony formation 8 weeks after treatment with busulfan, it can be concluded that only in 1 and 2 mg/kg of busulfan, recovery of endogenous germ cells was performed. In conclusion, the results demonstrated that intra-testicular injections of busulfan (2 mg/kg) reduced spermatocytes population in ram testis within 5 weeks of treatments, and this effect was reversible within 8 weeks of injection. However, it was not recommended to inject 4 mg/kg busulfan into the peritoneal cavity or testis of lambs based on its side effects.
Highlights
Transplantation of male animal germ cells is important because it is used to produce transgenic animals, propagation of elite sires, and conservation of genetic resources (Herrid et al, 2009). Using this technology in sheep industry is of great importance due to the problems associated with sperm cryopreservation, artificial insemination, and embryo transfer, which are used as a tool for transporting and disseminating genetics (RodriguezSosa et al, 2006)
Busulfan side effects In the first experiment, two days after intraperitoneal busulfan injection (4 mg/kg), two lambs died of diarrhea with lethargy and the other ones were injured on their mouth after 4 days and died after 20 days
The left testicular weight was significantly lower in the busulfan group than in the control group (p
Summary
Transplantation of male animal germ cells is important because it is used to produce transgenic animals, propagation of elite sires, and conservation of genetic resources (Herrid et al, 2009). The successful transplantation of spermatogonial stem cells (SSCs) largely depends on the preparation of the recipients, which destroys and suppresses internal germ cells and spermatogenesis, thereby enabling the replacement of transplanted SSCs and beginning donor-derived spermatogenesis (Ma et al, 2011) Various methods, such as genetically infertile hosts, chemotherapy, and radiation treatments, are used to eliminate host germ cells, leading to the improving the colonization of SSCs in the recipients (Lin et al, 2017).
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