Forums Index Forums Embryology (includes PGT & IVM) Sperm Cryopreservation and Post-Thaw Quality

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  • AvatarRyan Rose
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    Hello Embryology SIG,
    I thought I would start a new thread regarding sperm cryopreservation. We have several students this year working on optimisation of sperm cryopreservation techniques and it would be fantastic to get a worldwide opinion the subject.
    It is an interesting topic and quite overlooked in some regard. It’s overlooked because, although cryopreservation of sperm has been shown to cause morphological alterations and a significant decline in motility, viability, chromatin stability, and membrane integrity, it is thought that these deficits are solved by sperm selection techniques post-thaw (density gradient and swim-up being the most used techniques for sperm selection). In the event any doubt remains about the quality of the sample, convert the cycle to ICSI and the problem is solved…
    So our questions to this community would be;
    What techniques do you use for cryopreservation?
    Programmable freezing? Slow freezing? Rapid freezing? Vitrification?
    Many Australian ART clinics would use a combination of programmable freezing and slow freezing. However, it also seems to be due to a time factor. Some clinics state that programmable freezing post-thaw survival is comparable to “just leaving it in the -80 freezer over night followed by submersion in liquid nitrogen”.
    In these circumstances, it’s possible the method of post-thaw survival assessment would be a subjective difference. This leads me to my next question.
    How do you assess post-thaw quality?
    Many clinics do not have an operating andrology lab and either send semen samples to a third-party service provider for an assessment or simply check for number/motility by via a cell counter, Makler or similar device. Are any clinics running any of the DNA damage test routinely for cryopreserved samples, and if yes, why?
    What cryoprotectants do you use for sperm cryopreservation? Is every clinic using just glycerol, or at least part glycerol?
    I would confidently suggest most clinics in my region would be using glycerol or sucrose/glycerol mix. However, since the majority would be purchasing freezing medium, I think most commercially available medium would be a glycerol/glycerol mix? It would be interesting to hear if anyone makes their own medium, and why?
    I suspect differences in sperm cryopreservation practices will be region specific and if this is the case it would be an interesting observation itself. I think by openly discussing topics like this together it could open some interesting questions we could explore further. And at the very least our students and trainees will expand their knowledge.
    Happy sharing 😊

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