Cryopreservation of spermatozoa

Cryopreservation of spermatozoa (reprinted from Schenkel, Transgene Tiere 2. edition 2006, ISBN-10 3-540-28267-X, with friendly permit of Springer-Verlag, Heidelberg)
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Spermatozoa are prepared from the Vas deferens and the Epididymis of the male donors, high amounts of spermatozoa can be obtained. One donor might be sufficient for 10 to 15 parallel straws, 7 to 10 male donors are needed for the cryopreservation of a line. The samples are frozen in the steam of liquid nitrogen and afterwards plunged into LN2 permitting an unlimited storage.

© dkfz.de

In contrast to the relatively simple technique of cryopreservation an in vitro-fertlisation (IVF) is required following the revitalisation of spermatozoa. This technique exhibits varying success and is not available for all genetic backgrounds (Fig. 4). IVF needs a high number of oocyte donors. Following an over night culture 2-cell-embryos will be obtained and are subsequently subjected to an embryo transfer. In case of a general failure of the IVF alternative technologies are available. However, these techniques are very complex and cannot be applied -at least to date- as standard procedures.
To examine the quality of a frozen sample one of the parallel straws will be revitalized and the mobility and motility of the thawed spermatozoa will be examined using a dissection microscope.

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