Ohta Hiroshi, Sakaide Yuko, Wakayama Teruhiko
Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN, Kobe, Japan.
J Reprod Dev. 2008 Aug;54(4):295-8. doi: 10.1262/jrd.20027. Epub 2008 May 13.
We previously demonstrated that testicular spermatozoa can be preserved as frozen testicular sections, allowing us to preserve male gametes in less space than conventional methods. However, it remains unclear whether the testicular spermatozoa can be preserved for a long period using this procedure. In this study, we examined the function of testicular spermatozoa preserved as frozen testicular sections for l year at -30 or -80 C. Testicular spermatozoa were successfully retrieved from frozen testicular sections preserved at either -30 or -80 C, and their function was assessed using intracytoplasmic sperm injection (ICSI). Over 90% of the oocytes injected with long-term preserved testicular spermatozoa formed pronuclei, which was a frequency similar to that obtained with spermatozoa preserved for a short term, indicating that the testicular spermatozoa retained oocyte activation factor(s). Approximately 70% of the fertilized oocytes developed to 2-cell stage embryos, and 9.3 to 12.8% of the embryos developed to term after transfer into pseudopregnant females, regardless of the preservation temperatures examined. These results indicate that the birthrates of progeny did not differ between the preservation temperatures examined. They also indicate that male gametes can be preserved in testicular frozen sections for at least 1 year without loss of function.
我们之前证明，睾丸精子可以作为冷冻睾丸切片保存，这使我们能够以比传统方法更小的空间保存雄性配子。然而，使用这种方法睾丸精子是否能长期保存仍不清楚。在本研究中，我们检测了在-30℃或-80℃下作为冷冻睾丸切片保存1年的睾丸精子的功能。从保存在-30℃或-80℃的冷冻睾丸切片中成功获取了睾丸精子，并使用卵胞浆内单精子注射（ICSI）评估其功能。超过90%注射了长期保存的睾丸精子的卵母细胞形成了原核，这一频率与短期保存的精子相似，表明睾丸精子保留了卵母细胞激活因子。无论检测的保存温度如何，大约70%的受精卵发育到2细胞期胚胎，9.3%至12.8%的胚胎在移植到假孕雌性体内后发育至足月。这些结果表明，在所检测的保存温度下，后代的出生率没有差异。它们还表明，雄性配子可以在睾丸冷冻切片中保存至少1年而不丧失功能。
