小鼠未成熟睾丸同种异体移植的动物模型建立和改进

Establishment and improvement of animal model for allotransplantation of mouse immature testis

目的:初步探索采用同品系ICR小鼠作为受体小鼠,建立小鼠未成熟睾丸同种异体移植的动物模型。方法:采用6天龄ICR乳鼠未成熟睾丸作为供体,移植于已去势的同品系ICR受体小鼠颈背部皮下。移植后3、4、5周处死小鼠,观察移植物成活状况和移植物体积,HE染色评估移植物组织病理学的情况。荧光定量PCR检测异位成活的移植物组织内TESK1mRNA表达水平,供体睾丸和相应周龄小鼠睾丸组织作为对照。结果:移植后小鼠睾丸体积增长明显,于移植4周后进入平台期,移植前体积(17.3±2.6)mm3,移植3周组体积(203.1±14.3)mm3,移植4周组体积(236.4±27.5)mm3,移植5周组体积(220.9±17.4)mm3。组织病理学显示,移植物组织内曲细精管发育良好,生精功能正常,移植组曲细精管完整性和中空率3组间差异无统计学意义(P>0.05)。TESK1mRNA表达水平随着移植时间延长逐渐增高,移植前TESK1mRNA水平为对照组的(1.19±0.1)倍,移植3周组mRNA水平为移植前小鼠的(6.74±0.8)倍,移植5周组为移植3周组的(11.23±1.2)倍,差异有统计学意义(P<0.05)。结论:采用封闭系ICR小鼠作为受体的ICR小鼠未成熟睾丸移植是可行的。

Objective：To explore the immature testis allotransplantation model in the mouse using the same strain ICR mouse as a recipient,and to make a improvement based on the previous reports.Methods：The donor was the immature testis of six-day ICR-mouse.The recipient was the same strain ICR mouse.The immature testis of ICR mice were transplanted into the back of the neck in the recipient mice.After 3,4,5weeks,the status and the volume of the allograft were observed,and then the histopathology of the allograft was assessed.The level of TESK1 mRNA expression was analyzed by Q-PCR.Results：The volume of the allograft increased after allotransplantation.The average volume was（17.3±2.6）mm3 before transplantation,（203.1±14.3）mm3 after 3weeks of transplantation,（236.4±27.5）mm3 after 4weeks of transplantation,（220.9±17.4）mm3 after 4weeks of transplantation.Histopathologic results showed that seminiferous tubules developed well and the spermatogenic function recovered after immature testis allotransplantation.No significant differences of seminiferous tubules intact conditions were found among three groups after allotransplantation（P〉0.05）.The expression level of TESK1 mRNA increased after transplantation.The relative expression of TESK1 mRNA was（1.19±0.1）times before transplantation,（6.74±0.8）times in 3-week allograft,and（11.23±1.2）times in 5-week allograft the number of the control group（P〈0.05）.Conclusion：It is feasible that ICR mouse immature testis were allotransplanted using the same strain ICR mouse as a recipient.