Vitrified Semen Thawing at 100 Degree C. & 45 Degree C…

We are working on whole semen freezing by technique of Vitrification. Semen freezing by Vitrification is still under trial. Hence Vitrified Semen Thawing is least discussed topic. We freezed three cryovials of each semen sample.. One vial in each collection remain control sample. We freeze control sample by conventional slow freezing method. Two more cryovials we freezed of same semen samples by method of vitrification.

The aim of this study is to compare the quality of post thaw motility of vitrified semen: When thawed at different temperature.

  1. By using conventional method of thawing: control

    Cyrovial taken out from liquid nitrogen & kept in incubator at 330 C for 45 minutes.

  2. By thawing at 100 degree C (in boiling water)

    After taking out from liquid nitrogen, cryovial was waved in the air for few second and finally plunged in boiling water.

  3. By thawing at 45 degree C

    After taking out from liquid nitrogen, cryovial was dipped in water at 45O C

Material used: Post Wash CP to freeze one cryovial by conventional method., Pre Wash Vitri Media to vitrify semen sample, double walled cryovial, liquid nitrogen, etc.

Semen Vitrification:

After collection whole semen kept in incubator at 33 degree C for 45 minutes – till complete liquefaction.    

One ml of Pre Wash Vitri Media slowly added to completely liquefied and mixed semen. Mixed regularly and gently. Filled in cryovials. Kept at room temperature for 15 minutes. All cryovials plunged in liquid nitrogen.

After 24 hour we thawed all cryovials. Control cryovial was thawed as usual being done routinely. For vitrified semen we used two different protocol of thawing.

A simplest way to thaw vitrified semen

Conclusion: Vitrified Semen Thawing

We did not notice any gross difference in both method of thawing.  Thawing even at 100 degree C is equally good in case of vitrified semen.   We compared with conventional post thaw recovery of semen.  And our observation is relatively fast motility in vitrified sample. Which indicate somewhere that vitrified semen may give better result in term of conception

Moreover we understand, our study is limited to post thaw recovery of motility. Even sample size is small. But we can conclude with confidence that semen can be vitrified with successful post thaw recovery of motility.

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