Over the last forty years, in vitro fertilization, which has expanded to assisted reproductive technologies (ART), has gone from an experimental procedure to the mainstay of infertility treatment. A technique that onc...Over the last forty years, in vitro fertilization, which has expanded to assisted reproductive technologies (ART), has gone from an experimental procedure to the mainstay of infertility treatment. A technique that once made news with each birth is now responsible for 2% - 3% of the babies born in several nations of the world. This has happened due to significant advances in hormone therapies, culture techniques, and the specialization of equipment designed to support oocytes and embryos. However, for all the advances made to support female fertility, little has changed in male treatment since the advent of intracytoplasmic sperm injection in the early 1990’s. Recently, a number of authors have documented problems with sperm preparation techniques. Some report DNA damage, others membrane and organelle issues, all of which potentially hamper fertilization rates and possibly take-home baby rates. Further, as the clinical workload of ART has increased and staffing shortages have become critical, all labs are looking for simpler, more efficient ways to perform job functions. This study describes a simple, one-step method for preparing semen samples for ART. This new technique minimizes excessive manipulation of the sample compared to current standards and is less likely to cause cell damage. Preliminary results suggest a significant enhancement in recovered sample motility and an optimal sample for ART procedures with minimal sample manipulation.展开更多
Cryopreservation is currently the only effective tool for long-term storage of semen in most species. However, it is well-recognized that, even in species that freeze well, some individuals resist cryopreservation. Wo...Cryopreservation is currently the only effective tool for long-term storage of semen in most species. However, it is well-recognized that, even in species that freeze well, some individuals resist cryopreservation. Work from this laboratory has demonstrated a relationship between maternal lipid content and the chemical constitution of the embryos they produce. The objective of the present study was to determine if a similar relationship might exist in paternal body chemistry and the animal’s semen sample and if such a difference could be determined with a simple weight test. Semen samples were obtained from cattle with known differences in body composition. The samples first underwent semen analysis and were then prepared as either cell-free (CF) or neat specimens (NS). Known volumes of each sample were weighed, and the remainder of the samples was analyzed for lipids, total proteins, and total carbohydrates using a series of spectrophotometric assays and blood chemistry techniques. As expected, weight differences were seen in the CF vs NS preparations of individual semen samples (p < 0.001). Differences were also found in triglycerides (p < 0.001), glucose (p < 0.001), total protein (p < 0.001), and fructose (p < 0.009) of individuals with differing body composition. Statistical analysis suggested a non-linear correlation between the observed weights and total protein (p < 0.047) as well as triglyceride levels (p < 0.003). Together, these data suggest it might be possible to develop an algorithm to allow adjustment in cryoprotectants based on a simple weight procedure, allowing modification of cryoprotectants on an individual basis and potentially improving outcomes for valuable animals currently classified as “poor freezers”.展开更多
While generally recognized as a potential source of contamination during the collection process, lubricants are often used at the preference of the male partner to prevent irritation. While older lubricants have been ...While generally recognized as a potential source of contamination during the collection process, lubricants are often used at the preference of the male partner to prevent irritation. While older lubricants have been studied, there is currently no conscience within programs as to what constitutes a “safe” lubricant. The object of the current study was designed to evaluate chemically unique “next generation” lubricants in comparison to lubricants currently in use in fertility treatment and/or recognized as fertility safe;the first was a silicone-based lubricant, the second is a water-based, plant-based organic compound, in comparison to two established medical lubricants and a control. Twelve deidentified semen samples from the clinical andrology laboratory were used to test the lubricants following semen analysis. In order to enter the study, the sample had to have a minimum of 30 × 10<sup>6</sup> motile cells. Samples were then processed using a simple sperm wash modified to reconstitute the pellet into a final volume of 7 mL. Half-milliliter aliquots were then transferred into 13 wells of a standard 24-well culture plate. One well was used as a control. The remaining wells received one of the four lubricants at one of three volumes (10, 50, or 100 uL), producing 12 treatment combinations (four lubricants + 3 concentration levels) and the control. The samples were then cultured at room temperature for 24 hours. At times 0, 1, 3, 12, and 24 hrs, the plate was agitated to remix the sample, and a 4 uL aliquot of each well was analyzed for standard semen parameters using a computer-assisted semen analyzer. Results indicated the expected decrease in semen parameters over time in all treatments (P < 0.001). There was also a dose-dependent drop in most of the lubricants. However, samples contaminated with the newer lubricants appeared to maintain semen parameters similar to the controls at all but the 100 ul level of contamination, while the older lubricants caused decreases in sperm function at much lower concentrations. While semen parameters alone should not be the only criteria for the selection of a lubricant, the present study suggests newer formulations of lubricant are less likely to interfere with sperm function.展开更多
文摘Over the last forty years, in vitro fertilization, which has expanded to assisted reproductive technologies (ART), has gone from an experimental procedure to the mainstay of infertility treatment. A technique that once made news with each birth is now responsible for 2% - 3% of the babies born in several nations of the world. This has happened due to significant advances in hormone therapies, culture techniques, and the specialization of equipment designed to support oocytes and embryos. However, for all the advances made to support female fertility, little has changed in male treatment since the advent of intracytoplasmic sperm injection in the early 1990’s. Recently, a number of authors have documented problems with sperm preparation techniques. Some report DNA damage, others membrane and organelle issues, all of which potentially hamper fertilization rates and possibly take-home baby rates. Further, as the clinical workload of ART has increased and staffing shortages have become critical, all labs are looking for simpler, more efficient ways to perform job functions. This study describes a simple, one-step method for preparing semen samples for ART. This new technique minimizes excessive manipulation of the sample compared to current standards and is less likely to cause cell damage. Preliminary results suggest a significant enhancement in recovered sample motility and an optimal sample for ART procedures with minimal sample manipulation.
文摘Cryopreservation is currently the only effective tool for long-term storage of semen in most species. However, it is well-recognized that, even in species that freeze well, some individuals resist cryopreservation. Work from this laboratory has demonstrated a relationship between maternal lipid content and the chemical constitution of the embryos they produce. The objective of the present study was to determine if a similar relationship might exist in paternal body chemistry and the animal’s semen sample and if such a difference could be determined with a simple weight test. Semen samples were obtained from cattle with known differences in body composition. The samples first underwent semen analysis and were then prepared as either cell-free (CF) or neat specimens (NS). Known volumes of each sample were weighed, and the remainder of the samples was analyzed for lipids, total proteins, and total carbohydrates using a series of spectrophotometric assays and blood chemistry techniques. As expected, weight differences were seen in the CF vs NS preparations of individual semen samples (p < 0.001). Differences were also found in triglycerides (p < 0.001), glucose (p < 0.001), total protein (p < 0.001), and fructose (p < 0.009) of individuals with differing body composition. Statistical analysis suggested a non-linear correlation between the observed weights and total protein (p < 0.047) as well as triglyceride levels (p < 0.003). Together, these data suggest it might be possible to develop an algorithm to allow adjustment in cryoprotectants based on a simple weight procedure, allowing modification of cryoprotectants on an individual basis and potentially improving outcomes for valuable animals currently classified as “poor freezers”.
文摘While generally recognized as a potential source of contamination during the collection process, lubricants are often used at the preference of the male partner to prevent irritation. While older lubricants have been studied, there is currently no conscience within programs as to what constitutes a “safe” lubricant. The object of the current study was designed to evaluate chemically unique “next generation” lubricants in comparison to lubricants currently in use in fertility treatment and/or recognized as fertility safe;the first was a silicone-based lubricant, the second is a water-based, plant-based organic compound, in comparison to two established medical lubricants and a control. Twelve deidentified semen samples from the clinical andrology laboratory were used to test the lubricants following semen analysis. In order to enter the study, the sample had to have a minimum of 30 × 10<sup>6</sup> motile cells. Samples were then processed using a simple sperm wash modified to reconstitute the pellet into a final volume of 7 mL. Half-milliliter aliquots were then transferred into 13 wells of a standard 24-well culture plate. One well was used as a control. The remaining wells received one of the four lubricants at one of three volumes (10, 50, or 100 uL), producing 12 treatment combinations (four lubricants + 3 concentration levels) and the control. The samples were then cultured at room temperature for 24 hours. At times 0, 1, 3, 12, and 24 hrs, the plate was agitated to remix the sample, and a 4 uL aliquot of each well was analyzed for standard semen parameters using a computer-assisted semen analyzer. Results indicated the expected decrease in semen parameters over time in all treatments (P < 0.001). There was also a dose-dependent drop in most of the lubricants. However, samples contaminated with the newer lubricants appeared to maintain semen parameters similar to the controls at all but the 100 ul level of contamination, while the older lubricants caused decreases in sperm function at much lower concentrations. While semen parameters alone should not be the only criteria for the selection of a lubricant, the present study suggests newer formulations of lubricant are less likely to interfere with sperm function.
