The Effect Of Fertility And Physiological Function After Spermatogonial Stem Cells Transplantation Recipient Preparated With Busulfan In Mouse

Spermatogonial stem cells (SSCs) represent a crucial resource for male fertility and act as the groundwork of spermatogenesis. The transplantation of spermatogonial stem cells lay the foundation for the study of spermatogenesis, male fertility and new transgenic technology. To prepare recipient was an important part of the technology of spermatogonial transplantation. And it was also the premise and foundation and the necessary conditions of a successful transplantation. The efficiency of recipient preparation directly related to the effect of transplantation. Intra-peritoneal of busulfan is used more frequently as this method was more simple and convenient.But the operation created a considerable side effect and even caused death sometimes which influenced the clinically application. In this study,recipient was treated with testicular injection of busulfan, performed a blank determination with testicular injection of50%DMSO and a control with intra-peritoneal of busulfan. We measured fertility and testis physiological parameters (mortality,body and testis weights, hemogram, sperm counts, fertility and histological assessments of seminiferous tubules) and quantified the expression of glial-cell-line-derived neurotrophic factor (GDNF) transcripts at different time.There has no death after testicular injected,while the intra-peritoneal injected groups with mortalitys of18%、39.3%、42.7%, respectively. Through hemogram analysis, we found that there are significantly decline in RBC,HGB,WBC and PLT except MCHC after intra-peritoneal injected.In testicular injected groups, RBC、HGB、MCHC didn’t change significantly,but obvious decline was seen in WBC、PLT.And the decline is less than that of Intra-peritoneal injection,all items are quickly return to a normal state.40mg/kg and6、8、10mg/kg does can make a absolutely empty in seminiferous tubules. But4mg/kg does can not causes the same result.8mg/kg and10mg/kg dose also can result in an absolutely empty,while a great deal of testes are hardening and can’t recover at all. The variation tendency of sperm counts, relative testis weight and GDNF are consistent with that of seminiferous tubules. The results show that short-term infertility result from intra-peritoneal and testicular injection of busulfan,which can be recovered by themselves. And the recovery rate of mice fertility treated with testicular injection is quickly than that with intra-peritoneal injection.The sex ratio of the offspring did not change by the treatment,though it made an impact to fertility in a certain period of time.A conclusion was demonstrated in this study that the toxic action to animal by bilateral testicular injection of busulfan was significantly lower than that by intra-peritoneal injection, It can be used to preparate recipiention for SSCs transplantation by testicular injection of busulfan.