SD大鼠胞质内单精注射技术方法的建立

目的建立SD大鼠胞质内单精注射操作程序。方法和结果实验1：用直径为7～10μm和2-4μm的注射针以及相应的注射方法进行大鼠ICSI,ICSI后卵母细胞存活率（30．5％vs．61．3％）和卵裂率（12．5％vs．51．1％）均差异显著（P〈0．05）;实验2：分别在hCG后14h、16h和18h取卵进行ICSI,三组存活率（77．4％、74．1％vs．69．6％）差异不显著（P〉0．05）;14h和16h组的卵裂率（60．8％、56．0％vs．31．3％）与18h组差异显著（P〈0．05）;实验3：用不同的显微操作液H-mR1CM和H-mKRB进行大鼠ICSI,结果卵存活率相近（79．2％vs．75．9％）,卵裂率（70．5％vs．74．7％）差异不显著（P〉0．05）,但8-细胞发育率（43．5％vs．61．9％）差异显著（P〈0．05）,囊胚发育率差异极显著（P〈0．01）。结论大鼠ICSI时在注射hCG后14～16h取卵最佳,采用2—4pan直径的注射针、H—mKRB作为操作液更有利于卵的发育。

Establishment of Introcytoplasmic Sperm Injection Technology in SD Rat

Objective To establish the procedure of ICSI with Piezo-driven micromanipulator in SD rat. Methods and Results Experiment 1 ： Intracytoplasmic injection of rat sperm heads was performed using 7 - 10 μm and 2 - 4 μm diameter pipettes and different injected method respectively. The survived rate is 30.5% and 61.3%, the cleaved rate is 12.5% and 51.1% . Experiment 2 ： Oocytes were collected at 14h, 16h, 18h after given hCG, the ICSI survived rate is 77.4 %, 74.1% and 69.6 %, cleaved rate were 60.8 % ,56.0 % and 31.3 % respectively. Experiment 3 ： Injected the sperm heads in H-mR1CM and H-mKRB operation medium, the survived rate is 79.2% and 75.9%, the cleaved rate is 70.5% and 74.7%, oocyte-fertilized were further cultured in vitro, the 8-cell rate is 43.5 % and 61.9 %, the blastocyst rate is 21.2 % and 35.1%. Conclusion The results indicate that oocytes collected at 14- 16 h after given hCG, using the small-bore pipette and H-mKRB is propitious to rat ICSI and the development of ICSI embryos.