Viable rat offspring derived from oocytes intracytoplasmically injected with freeze-dried sperm heads
Viable rat offspring derived from oocytes intracytoplasmically injected with freeze-dried sperm heads
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DOI：
10.1017/s096719940500300x
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发表时间：
2005-02-01
期刊：
影响因子：
1.7
通讯作者：
Hochi, S
中科院分区：
文献类型：
--
作者：
Hirabayashi, M;Kato, M;Hochi, S
本研究采用2 × 3析因设计实验，研究大鼠冷冻干燥精子在卵胞浆内单精子注射（ICSI）后能否参与足月发育。用超声处理和非超声处理制备SD大鼠附睾尾部精子悬液。将超声处理和非超声处理的精子悬液进行冻融处理（FT组; 100 μ l样品在液氮蒸气中冷却，在196 ℃保存1天，并在25 ℃水浴中解冻）和冷冻干燥（FD组;将100 μ l样品在液氮中冷冻20秒，冻干6小时，在4 ℃下储存2天，并用100 μ l超纯水再水化），或立即用于ICSI（新鲜对照组）。使用直径为2-4 μ m的压电驱动微量移液管将精子头显微注射到裸露的SD卵母细胞中。将假定的受精卵转移到假孕Wistar雌性大鼠的输卵管中。从所有六个实验组中产生有活力的大鼠后代。大鼠精子的超声处理。结果表明，F1组、FT组、FD组的子代率分别为23.3%、35.0%、9.2%和2.5%。顶体区域似乎是完整的，即使在超声波FT和FD治疗以及在新鲜的控制，而精子膜的外侧背侧区域或多或少损坏的超声波处理，FT和FD样品。因此，冷冻干燥的精子成功地参与了足月发育，通过在大鼠中应用ICSI证明。
A 2 x 3 factorial designed experiment was conducted in order to examine whether freeze-dried rat spermatozoa can participate in full-term development following intracytoplasmic sperm injection (ICSI). A sperm suspension from cauda epididymides of Sprague-Dawley (SD) rats was prepared with or without ultrasonic treatment. The sonicated and non-sonicated sperm suspensions were processed for freeze-thawing (FT groups; 100 mu l sample was cooled in liquid nitrogen vapor, stored for 1 day at 196 degrees C, and thawed in a 25 degrees C water bath) and freeze-drying (FD groups; 100 mu l sample was frozen in liquid nitrogen for 20s, lyophilized for 6 h, stored at 4 degrees C for 2 days, and rehydrated with 100 mu l ultrapure water), or were subjected to immediate use for ICSI (fresh control groups). The sperm heads were microinjected into denuded SD oocytes using a piezo-driven micropipette 2-4 mu m in diameter. The presumptive zygotes were transferred into oviducts of pseudopregnant Wistar female rats. Viable rat offspring were produced from all six experimental groups. Ultrasonic treatment of rat spermatozoa. was effective in increasing the offspring rate (23.3% vs 6.7% in fresh control groups, 35.0% vs 7.6% in FT groups, 9.2% vs 2.5% in FD groups). The acrosomal region appeared to be intact even after ultrasonic FT and FD treatments as well as in the fresh controls, while the lateral dorsal region of the sperm membrane was more or less damaged in the sonicated, FT and FD samples. Thus, the successful participation of freeze-dried spermatozoa in full-term development was demonstrated by applying ICSI in the rat.