Burgstaller Sophie M, Auer Kerstin E, Rülicke Thomas
Department of Biomedical Sciences and Pathobiology, University of Veterinary Medicine Vienna, Vienna, Austria.
Ludwig Boltzmann Institute for Hematology and Oncology, Vienna, Austria.
J Assist Reprod Genet. 2024 Sep;41(9):2537-2546. doi: 10.1007/s10815-024-03201-x. Epub 2024 Jul 17.
Mouse spermatozoa for archiving laboratory mice or for in vitro fertilization (IVF) are routinely obtained from the cauda epididymis of adult males sacrificed for this purpose. To avoid the death of the donor, we tested whether a precisely timed interruption of the mating act could be used for repeated sperm collection from laboratory mice.
Sperm donors (B6D2F1) were mated with a receptive female, and mating behavior was observed. The stud was separated from the female 1-2 s after the onset of the ejaculatory shudder. The ejected copulatory plug with the yellowish viscous ejaculate was carefully removed from the penile cup.
A total of 80 ejaculates were successfully obtained from 100 ejaculations. The latency to first mount was 1.1 ± 1.1 min (mean ± SD) and to ejaculation 8.1 ± 4.7 min. The average number of mounts to ejaculation was 10.5 ± 5.8, and the mean number of spermatozoa per collected ejaculate was 1.86 ± 1.05 × 10. An average fertilization rate of 76% was observed after IVF.
Separating the stud from the female just before ejaculation is feasible, easy to learn, and requires no special equipment. The sperm count of collected ejaculates is lower than natural ejaculations, but higher than previous in vivo sperm collection methods achieved. We recommend this simple sperm collection method in mice, especially when the donor cannot be sacrificed and/or repeated sperm collection from the same animal is required for experimental purposes.
用于存档实验小鼠或体外受精（IVF）的小鼠精子通常从为此目的而处死的成年雄性小鼠的附睾尾部获取。为避免供体死亡，我们测试了是否可以通过精确计时中断交配行为来从实验小鼠重复采集精子。
将精子供体（B6D2F1）与处于接受状态的雌性小鼠交配，并观察交配行为。在射精颤抖开始后1 - 2秒将雄性小鼠与雌性小鼠分开。小心地从阴茎套中取出带有淡黄色粘性精液的射出的交配栓。
从100次射精中成功获得了80次射精。首次爬跨的潜伏期为1.1±1.1分钟（平均值±标准差），射精潜伏期为8.1±4.7分钟。每次射精的平均爬跨次数为10.5±5.8次，每次采集的射精中精子的平均数量为1.86±1.05×10个。体外受精后观察到平均受精率为76%。
在射精前将雄性小鼠与雌性小鼠分开是可行的，易于掌握，且无需特殊设备。采集的射精中的精子数量低于自然射精，但高于以往的体内精子采集方法所获得的数量。我们推荐这种简单的小鼠精子采集方法，特别是当不能处死供体和/或出于实验目的需要从同一动物重复采集精子时。
