Bouamama N, Briot P, Testart J
Cadi Ayyad university of, Marrakech, Maroc.
Gynecol Obstet Fertil. 2003 Feb;31(2):132-5. doi: 10.1016/s1297-9589(03)00003-1.
In case of severe oligozoospermia or following testicular biopsy in case of azoospermia, fertilization remains possible by intracytoplasmic sperm injection (ICSI). However, if there are only a few spermatozoa available to be preserved for a delayed ICSI, these gametes must be cryopreserved according to a specific method. We have found that, using the straw classical technique, less than 2 sperm were recovered at thawing when 20 sperm were frozen and no sperm were available following freezing-thawing of only 1 to 10 sperm per straw. Then, the feasibility of programmed ICSI is always uncertain in these cases and unnecessary medical acts (ovarian stimulation and egg collection) as well as patient disappointment are frequent.
We have tested a new technique by introducing (with an ICSI pipette) 1 to 100 sperm in a microdrop (0.5 microl) of freezing medium setting in a culture dish under paraffin oil.
Following freezing-thawing the dish, all (100%) of the frozen spermatozoa were recovered.
Since only motile sperm were frozen, the sperm which are non motile after thawing (more than 50%) are nevertheless considered usable for ICSI.
在严重少精子症的情况下,或者在无精子症患者进行睾丸活检后,通过卵胞浆内单精子注射(ICSI)仍有可能实现受精。然而,如果仅有少量精子可供保存用于延迟的ICSI,则必须按照特定方法对这些配子进行冷冻保存。我们发现,采用传统的细管技术,当冷冻20个精子时,解冻后回收的精子少于2个;而每根细管仅冷冻1至10个精子时,冻融后则没有可用的精子。因此,在这些情况下,程序化ICSI的可行性始终不确定,不必要的医疗行为(卵巢刺激和取卵)以及患者的失望情绪屡见不鲜。
我们测试了一种新技术,即将1至100个精子(使用ICSI移液管)引入置于石蜡油下培养皿中的一滴冷冻培养基(0.5微升)中。
对培养皿进行冻融后,所有(100%)冷冻的精子均被回收。
由于仅冷冻了活动精子,解冻后不活动的精子(超过50%)仍被认为可用于ICSI。