Effective production of recipient male pigs for spermatogonial stem cell transplantation by intratesticular injection with busulfan

Lin, Zhuoheng; Bao, Jiajing; Kong, Qingming; Bai, Yinshan; Luo, Fanghong; Songyang, Zhou; Wu, Yan; Huang, Junjiu

doi:10.1016/j.theriogenology.2016.10.021

Cited by 10 publications

(5 citation statements)

References 32 publications

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“…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. Lin et al (2017) found the recovery of endogenous germ cells slightly after three months of busulfan treatment in Seghers pigs (recovery of Pgp9.5 mRNA level).…”

Section: Discussionmentioning

confidence: 87%

“…In the first experiment, the lambs were injected intraperitoneal with 4 mg/kg doses of the dissolved busulfan in 40 mg/mL dimethyl sulfoxide (DMSO). In the second experiment, the direct injection of 1, 2 and 4 mg/kg doses of the dissolved busulfan in 40 mg/mL DMSO into both sides of the scrotums of left testis was conducted equally by using 5-mL syringes, as described by Lin et al, 2017. The 0 mg/kg group was considered as the control group, and the dose of the injected DMSO was the same as the experimental groups.…”

Section: Methodsmentioning

confidence: 99%

“…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).…”

Section: Introductionmentioning

confidence: 99%

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Spermatogonial stem cell survival in ram lambs following busulfan treatment

et al. 2020

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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.

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Section: Discussionmentioning

confidence: 87%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Spermatogonial stem cell survival in ram lambs following busulfan treatment

et al. 2020

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“…Consequently, busulfan treatment is the most common method used to prepare recipients of spermatogonial stem cells transplantation [27,28] and to study spermatogonial stem cells kinetics and fertility recovery [29,30,31]. Busulfan can eliminate almost all endogenous germ cells in the recipient, creating an empty space in the spermatogonial stem cells niche; therefore, this drug has been used successfully to prepare recipients in mice [32,33], rats [34,35], pigs [36,37], dogs [38], hamsters [39], monkeys [40], and roosters [41].…”

Section: Chemotherapymentioning

confidence: 99%

Effects of Chemotherapy and Radiotherapy on Spermatogenesis: The Role of Testicular Immunology

Itoh

Sakabe

2019

IJMS

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Substantial improvements in cancer treatment have resulted in longer survival and increased quality of life in cancer survivors with minimized long-term toxicity. However, infertility and gonadal dysfunction continue to be recognized as adverse effects of anticancer therapy. In particular, alkylating agents and irradiation induce testicular damage that results in prolonged azoospermia. Although damage to and recovery of spermatogenesis after cancer treatment have been extensively studied, there is little information regarding the role of differences in testicular immunology in cancer treatment-induced male infertility. In this review, we briefly summarize available rodent and human data on immunological differences in chemotherapy or radiotherapy.

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“…Qin et al (2016a) established a method of intratesticular injection of busulfan in mice for the preparation of recipients, and the high mortality rate was effectively overcome. Moreover, studies have shown that this intratesticular injection method has a large potential to improve recipient transplantation success in both pigs and mice (Ganguli et al 2016;Lin et al 2017). However, under physiological conditions, tight junctions and reactions between Sertoli cells (SCs) and SSCs help to stabilize the microenvironment inside the testicles, which ensures normal physiological functions of SSCs, such as proliferation, differentiation, and apoptosis.…”

Section: Introductionmentioning

confidence: 99%

Progress on the study of the mechanism of busulfan cytotoxicity

2018

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The preparation of spermatogonial stem cell (SSC) transplant recipients laid the technical foundation for SSC transplant technology and the understanding of spermatogenesis mechanisms. Busulfan is commonly used to prepare recipients for mouse SSC transplantation; however, its safety and efficiency have been questioned. This review summarizes the relationship between SSCs and Sertoli cells (SCs), and the mechanism of busulfan toxicity against sperm cells. We concluded that the proliferation, differentiation, and apoptosis of SSCs are regulated by SCs. The endogenous spermatogenic cells are depleted by busulfan treatment via alkylation of DNA, destruction of vimentin filament distribution, disruption of SSC differentiation, promotion of SSC dormancy, and generation of oxidative stress. However, the mechanisms require further exploration. The recent establishment of a model in vitro culture system has provided a good technical foundation to further explore these mechanisms, which will help us to find more efficient methods of recipient preparation and optimal transplantation times.

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Effective production of recipient male pigs for spermatogonial stem cell transplantation by intratesticular injection with busulfan

Cited by 10 publications

References 32 publications

Spermatogonial stem cell survival in ram lambs following busulfan treatment

Spermatogonial stem cell survival in ram lambs following busulfan treatment

Effects of Chemotherapy and Radiotherapy on Spermatogenesis: The Role of Testicular Immunology

Progress on the study of the mechanism of busulfan cytotoxicity

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