ABSTRACT
Successful sperm cryopreservation has been reported with methods that use methanol and powdered milk as cryoprotectants and microcapillaries as cryovessels[25]. However, our laboratory and others have observed inconsistent results with these methods[7,8]. Furthermore, the reported techniques can be inconvenient because they require complex manipulations of small volumes and are not compatible with largescale protocols. By performing a broad evaluation of cryoprotectants and sperm diluents and investigating the effect of increasing the volume of freezing medium on fertilization capacity, we developed a method that provides efficient recovery and an increased number of archived samples. We found that cryopreserving homogenized, whole testes in 10% N,N-dimethylacetamide (DMA) diluted in Buffered Sperm Motility Inhibiting Solution (BSMIS) permits efficient embryo recovery from four, 50 μl frozen samples [7,8].
