For instance, the formation of intracellular ice crystals and osmotic imbalance can damage the integrity of spermatozoa, membrane structure and function. Because the spermatozoa are highly differentiated, are very structural and are functionally motile, these kinds of damage results in the loss of viability, motility and fertility of spermatozoa. ĭespite the aforementioned advantages of cryopreservation, its process may directly or indirectly affect severe and irreversible damages to the spermatozoa with respect to the thermal shock (cold shock), formation of intracellular ice crystals, osmotic shock, stress of cryoprotectants and generator of reactive oxygen species (ROS). Furthermore, canine spermatozoa cryopreservation can be done even post mortem if the male dog dies accidently or unexpectedly, the procedures with collection of spermatozoa from the epididymis and cryopreservation are feasible to produce his offspring in the future via the artificial insemination. In addition, transportation of cryopreserved spermatozoa can make female dogs fertilized at a distant place, which can reduce their stress and shipping cost. Canine spermatozoa cryopreservation may help to create progeny of rare dog breeds, genetically valuable dogs such as the guide dog, detection dog and disease model dog, and the dog whose the owner wants to breed cryopreservation technology has contributed to the protocol of genome resource banking in canine as well. Of particular, there are additional benefits of spermatozoa cryopreservation in canine. Since the introduction of glycerol and dimethyl sulfoxide (DMSO) as the cryoprotectants, cryopreservation techniques have been extensively applied in many cells including spermatozoa (semen) spermatozoa cryopreservation is a way for preserving genetic information and diversity in several species for an unlimited time theoretically and enables male germplasm to use in the future. Keywords: Normal reference data, Canine spermatozoa, Fresh spermatozoa, Cryopreserved Spermatozoa, CryopreservationĬryopreservation indicates long-term storage in condition of subzero ℃ temperatures without the metabolic activity on the cellular systems. We hope that this review contributes to the advancement of cryobiology in canine spermatozoa. Here, we reviewed the recent articles to introduce various analytical methods for the canine spermatozoa as well as to establish the normal reference data for each analytical method in the fresh or cryopreserved canine spermatozoa, based on the results of the previous articles. Because the study designs with certain application of analytical methods are selective and varied depending on each experimental objective and laboratory condition, it is necessary to establish the normal reference data of the fresh or cryopreserved canine spermatozoa for each analytical method to monitor experimental procedure, to translate raw data and to discuss results. According as a number of researches have been conducted to overcome these problems and to advance cryopreservation technique, several analytical methods have been employed to evaluate the quality of the fresh or cryopreserved canine spermatozoa in regards to the motility, morphology, integrity of membrane and DNA, mitochondrial activity, ROS generation, binding affinity to oocytes, in vitro fertilization potential and fertility potential by artificial insemination. However, severe and irreversible damages to the spermatozoa during cryopreservation procedures such as the thermal shock (cold shock), formation of intracellular ice crystals, osmotic shock, stress of cryoprotectants and generator of reactive oxygen species (ROS) have been addressed. Especially, the canine spermatozoa cryopreservation has contributed on generation of progeny of rare/genetically valuable dog breeds, genome resource banking and transportation of male germplasm at a distant place. The cryopreservation has been extensively applied in many cells including spermatozoa (semen) during past several decades.
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