Akash Verma: Sperm Cryopreservation with FertiPro SpermFreeze
Akash Verma, Embryologist at Suman Hospital Ludhiana, shared on LinkedIn:
”Sperm Cryopreservation with FertiPro SpermFreeze
From Liquefaction to Post-Thaw Washing
Sperm cryopreservation is more than simply freezing a semen sample. Every stage-from liquefaction and cryoprotectant equilibration to thawing and post-thaw washing-can influence sperm recovery and motility.
SOP-Style Workflow
Liquefaction
Allow semen to liquefy at room temperature for 30 minutes.
Pre-freeze assessment
Assess volume, concentration, motility, and other required semen parameters.
Cryoprotectant addition
For FertiPro SpermFreeze:
1 mL semen + 0.7–1.0 mL SpermFreeze
Add the medium dropwise while gently swirling. The medium should be at room temperature to minimize cold shock.
Equilibration
Allow the mixture to equilibrate for 10 minutes at room temperature.
Straw loading
Load the mixture into labelled freezing straws, leaving approximately 1.5 cm air space at the end, then seal.
Freezing
Place the straws in the LN₂ vapour phase for at least 15 minutes, followed by rapid transfer into liquid nitrogen for storage at approximately −196°C.
Thawing
Place the required straw in tap water for 5 minutes at room temperature or 37°C.
Post-thaw dilution and washing
Recover the sample and dilute with suitable insemination medium, using at least 3 mL medium per 0.5 mL semen.
Centrifuge at:
300–350 × g – 15 minutes
Remove the supernatant and resuspend the sperm pellet in suitable medium.
Post-thaw assessment
Evaluate sperm concentration, motility and recovery according to the laboratory’s validated SOP.
Important laboratory point
For FertiPro SpermFreeze, the manufacturer states that cryopreservation is ideally performed on native semen rather than routinely washing before freezing. After thawing, the sample should be washed before further use such as IUI to remove the cryopreservation medium.
Cryopreservation principle:
Protect – Equilibrate – Freeze – Store – Thaw – Wash – Assess
Always follow the current manufacturer’s IFU and your laboratory’s validated SOP.”
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