單獨使用0.125 M trehalose 海藻糖 或 sucrose即可用於冷凍精蟲達69%存活率
兩段式(液態氮蒸氣>液態氮)冷凍精蟲優於直接至液態氮

Cryobiology. 2016 Oct;73(2):162-7. doi: 10.1016/j.cryobiol.2016.08.004. Epub 2016 Aug 4.

Cryopreservation of human spermatozoa with minimal non-permeable cryoprotectant.

Liu J1, Tanrikut C2, Wright DL3, Lee GY1, Toner M1, Biggers JD4, Toth TL5.

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Abstract

Cryopreservation of human spermatozoa is a commonly used technique in assisted reproduction, however freezing low concentrations of sperm while maintaining adequate post-thaw motility remains a challenge. In an effort to optimize post-thaw motility yields, low volumes of human sperm were frozen in polyimide-coated fused silica micro-capillaries using 0.065 M, 0.125 M, 0.25 M, or 0.5 M trehalose as the only cryoprotectant. Micro-capillaries were either initially incubated in liquid nitrogen vapor before plunging into liquid nitrogen, or directly plunged into liquid nitrogen. Post thaw sperm counts and motility were estimated. Spermatozoa that were initially incubated in liquid nitrogen vapor had greater post thaw motility than those plunged immediately into liquid nitrogen independent of trehalose concentration. The protective effect of 0.125 M d-glucose, 3-O-methyl-d-glucopyranose, trehalose, sucrose, raffinose, or stachyose were evaluated individually. Trehalose and sucrose were the most effective cryoprotectants, recovering 69.0% and 68.9% of initial sperm motility, respectively.