Cryopreservation of Surgically Retrieved Spermatozoa 手术所取精子的冷冻方法

With the new advances in cryotechnology, the cryopreservation of surgically retrieved spermatozoa has become a very appealing and occasionally indispensable approach for managing infertility due to severe male factor. There are some notable advantages in the efficient cryopreservation of a small amount of sperm.Specifically, it decreases the amount of surgical interventions needed so that expenses and risks are also reduced. Moreover, the use of cryopreserved sperm removes the issue of planning egg retrieval to coincide with a urologist’s schedule for the surgical retrieval of sperm. Moreover,performing the sperm retrieval prior to initiation of the in vitro fertilization (IVF) stimulation cycle eliminates the risk of no sperm being found on the day of egg retrieval.

随着冷冻技术取得新的进展，对于因严重男性因素导致的不孕症的案例，冷冻保存通过手术取出的精子，已经成为一种非常有吸引力且有时必不可少的方法。少量精子的有效冷冻保存有一些显著的优势。具体而言，它减少了手术所需的干预措施，从而降低了费用和风险。 此外，使用冷冻保存的精子，免去了因配合泌尿科医生的时间安排而协调取卵时间的麻烦。 此外，在试管促排周期开始之前安排取精手术，避免取卵当天无精子可用的风险。

Sperm extraction procedures are usually performed by aurologist. Subsequently, the retrieved sperm are transferred to the laboratory for assessment and freezing. For cryopreservation, epididymal and testicular sperm samples are diluted 1: 1 with test yolk buffer-glycerol cryoprotectant and aliquotted into cryovials. Vials are vapor frozen for 30 minutes prior to immersion in LN2. It is also helpful to freeze a separate 25 μL “test” aliquot for post-thaw evaluation. Before IVF cycle initiation, the “test aliquot” is thawed and assessed. This process allows patients to be adequately counseled before cycle initiation. Plans could be made for back-up donor specimens, fresh sperm harvest and oocyte freezing, in the event that the laboratory cannot locate motile sperm after extensive searching.

取精手术通常由泌尿科医生进行操作。 随后，取出的精子被转移到实验室进行评估和冷冻。对于冷冻保存，附睾和睾丸取出的精子样本用测试卵黄缓冲液-甘油冷冻保护剂 1:1 稀释并分装到冷冻管中。 放入液氮箱之前，将装有精液的试管蒸汽冷冻 30 分钟。 单独冻结一份25 μL的“测试”分试样，对于进行解冻精液后的评估也很有帮助。 在试管周期开始之前，“测试分试样”被解冻并用于评估精液指标。这一过程使患者在周期开始前有充分的咨询时间，以防实验室在精液样本中无法找到有活力的精子用于受精，患者可提前做好如下准备：购买他人捐赠的精液样本，或在取卵当天收集新鲜精液，又或把取出的卵子进行冷冻。