Zhang Wei, Xia Xiao-Yu, Ding Hui, Liu Ping, Cai Heng, Li Peng, Xu Chen, Li Zheng
Department of Urology/Center for Clinical Research of Stem Cells, Renji Hospital, Shanghai 200001, China.
Zhonghua Nan Ke Xue. 2009 Aug;15(8):703-7.
To establish a stable recipient mouse model for stem cell transplantation into seminiferous tubules and improve the traditional techniques for transplantation.
Sixty male ICR mice were equally divided into Groups A, B, C and D, and injected with Busulfan at 15 mg/kg, 30 mg/kg, 40 mg/kg and 0 mg/kg, respectively. The survival rate was recorded every day, and the testis weight and spermatogenesis of testicular tubules were determined at 4, 8 and 12 weeks after the injection. We improved the stem cell transplantation technique and designed a new transplantation device, which connected the nozzle end, syringe and puncture needle by a three-way joint. The nozzle end was used for tentative injection, and the syringe for drawing and then injecting the cell suspension.
Only one mouse in Group C died after the injection. At 4 weeks after Busulfan treatment, the testis weight decreased apparently in Groups A, B and C, with significant differences from D (P < 0.05). The differences remained significant at 8 weeks (P < 0.05), except between Groups A and D (P > 0.05), but at 12 weeks none of the first three groups showed any significant difference from Group D (P > 0.05). At 4 and 8 weeks, the rate of hollow seminiferous tubules was < 50% in Group A and > 50% in Groups B and C, and almost returned to normal at 12 weeks, with no significant differences among the three groups (P > 0.05), but it remained unchanged in Group D. The improved transplantation device increased the success rate (> 90%), lowered the donor cell loss (< 50 microl cell suspension needed for both testes) and shortened the process ( < 10 min for one testis).
Intraperitoneal injection of Busulfan at 30 mg/kg is suitable for the establishment of the recipient mouse model of stem cell transplantation. The improved transplantation device and methods help promote the efficiency and success rate of the transplantation operation.
建立一种稳定的将干细胞移植到生精小管的受体小鼠模型，并改进传统的移植技术。
将60只雄性ICR小鼠平均分为A、B、C、D四组，分别以15mg/kg、30mg/kg、40mg/kg和0mg/kg的剂量注射白消安。每天记录存活率，并在注射后4周、8周和12周测定睾丸重量和曲细精管的生精情况。我们改进了干细胞移植技术，设计了一种新的移植装置，该装置通过三通接头连接喷嘴端、注射器和穿刺针。喷嘴端用于试探性注射，注射器用于吸取并注射细胞悬液。
注射后C组仅1只小鼠死亡。白消安处理后4周，A、B、C组睾丸重量明显下降，与D组相比差异有统计学意义（P<0.05）。8周时差异仍有统计学意义（P<0.05），A组和D组除外（P>0.05），但12周时前三组与D组相比均无显著差异（P>0.05）。4周和8周时，A组中空曲细精管的比例<50%，B组和C组>50%，12周时几乎恢复正常，三组间差异无统计学意义（P>0.05），但D组保持不变。改进后的移植装置提高了成功率（>90%），降低了供体细胞损失（双侧睾丸所需细胞悬液<50μl），缩短了操作过程（单只睾丸<10分钟）。
腹腔注射30mg/kg白消安适合建立干细胞移植的受体小鼠模型。改进后的移植装置和方法有助于提高移植手术的效率和成功率。
