We have tested a variety of cryoprotectants for their efficacy in preserving viable sperm. Our best results were achieved from the following formula:
Separate the yolk of a chicken egg from the white. The yolk was then transferred into a 50 mL conical tube and diluted 1:1 with sterile water. (An egg yolk from a large egg is approximately 15mL). The contents were lightly vortexed to ensure homogenization.
The diluted egg yolk was then diluted 20% (v:v) in a solution consisting of 0.4 M sucrose and 20 mM sodium bicarbonate (sterile filtered).
The solution was then centrifuged at 13,000 rpm in a refrigerated centrifuge and the supernatant aliquotted as the cryoprotectant. Aliquots were stored at –80oC and thawed on ice when needed for use.
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Last update: Feb. 13, 2008