Controlled ice nucleation--Is it really needed for large-volume sperm cryopreservation?

Abstract

Controlled ice nucleation (CIN) is an integral stage of slow freezing process when relatively large volumes (usually 1 mL or larger) of biological samples in suspension are involved. Without it, a sample will supercool to way below its melting point before ice crystals start forming, resulting in multiple damaging processes. In this study, we tested the hypothesis that when freezing large volumes by the directional freezing technique, a CIN stage is not needed. Semen samples collected from ten bulls were frozen in 2.5-mL HollowTubes in a split-sample manner with and without a CIN stage. Thawed samples were evaluated for viability, acrosome integrity, rate of normal morphology, and, using com..