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Cryorecovery of Mouse Sperm by Different IVF Methods Using MBCD and GSH.

Li MW, Glass OC, Zarrabi J, Baker LN, Lloyd KC - J Fertili In Vitro (2016)

Bottom Line: Different protocols incorporating methyl-β-cyclodextrin (MBCD) and reduced glutathione (GSH) have been reported to improve IVF recovery of cryopreserved mouse sperm on a C57BL/6 (J and N) genetic background.Therefore, in the present study we correlated sperm motility with fertilization rate and compared the efficiency of different IVF methods using various sperm samples so as to establish general guidelines for mouse sperm cryorecovery by IVF.Moreover, the presence of a large number of immotile sperm in the sperm-oocyte co-incubation drop was found to reduce IVF success which could be partially reversed by supplementation using monothioglycerol (MTG) during centrifugation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Mouse Biology Program, University of California, Davis, CA 95618, United States.

ABSTRACT

Different protocols incorporating methyl-β-cyclodextrin (MBCD) and reduced glutathione (GSH) have been reported to improve IVF recovery of cryopreserved mouse sperm on a C57BL/6 (J and N) genetic background. However, it is not clear which IVF protocol is most appropriate when using the various methods to cryorecover sperm with different sperm quality and sample volumes. Therefore, in the present study we correlated sperm motility with fertilization rate and compared the efficiency of different IVF methods using various sperm samples so as to establish general guidelines for mouse sperm cryorecovery by IVF. High linear correlation between sperm fertilization rate and progressive motility was found, R(2) was 0.9623 and 0.9993 for pre-freezing and post-thaw progressive motility, respectively. High amounts of cryoprotective agent (CPA) were observed to impair both sperm capacitation and fertilization. Moreover, the presence of a large number of immotile sperm in the sperm-oocyte co-incubation drop was found to reduce IVF success which could be partially reversed by supplementation using monothioglycerol (MTG) during centrifugation. It was concluded that the efficiency of IVF using cryorecovered mouse sperm in media containing MBCD and GSH can be predicted from sperm progressive motility. High concentrations of CPA and immotile sperm should be mitigated prior to IVF. The optimum IVF method should be selected based on sperm sample volume and sperm parameters.

No MeSH data available.


Related in: MedlinePlus

Linear regression between the mean of sperm progressive motility (pre-freeze and post-thaw) and the mean of fertilization rates of MBCD-GSH IVF. Ypre = pre-freeze progressive motility; Ypost = post-thaw progressive motility; X = IVF rate.
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Figure 2: Linear regression between the mean of sperm progressive motility (pre-freeze and post-thaw) and the mean of fertilization rates of MBCD-GSH IVF. Ypre = pre-freeze progressive motility; Ypost = post-thaw progressive motility; X = IVF rate.

Mentions: Sperm samples of different mouse lines had different sperm motility. To determine the correlation between sperm motility and their fertilization abilities, MBCD-GSH IVF procedures were performed using cryopreserved sperm samples from 46 knockout mouse lines on a C57BL/6N genetic background. Although there was a large degree of variation in the IVF rate of thawed mutant sperm samples for each progressive motility (pre-freeze or post-thaw), there was a significant linear correlation between the mean of the IVF rate and the mean of the progressive motility after the fertilization rates of the 46 IVF procedures (1 IVF procedure per mouse line using sperm of a heterozygous male) were sorted and separated into 5 categories: 5–19%, 20–29%, 30–39%, 40–59%, and 60–80% (Figure 1). The R2 were 0.9623 and 0.9993 for pre-freeze and post-thaw progressive motility, respectively (Figure 2). The higher the mean of the progressive motility (especially the post-thaw progressive motility), the higher the mean of the IVF rate. IVF rates were usually less than 30% when post-thaw progressive sperm motility was less than 10%. The linear equations for the two variables (X, IVF rate; Y, pre-freeze or post-thaw progressive motility) containing slopes and Y-intercepts when X=0 are shown in Figure 2.


Cryorecovery of Mouse Sperm by Different IVF Methods Using MBCD and GSH.

Li MW, Glass OC, Zarrabi J, Baker LN, Lloyd KC - J Fertili In Vitro (2016)

Linear regression between the mean of sperm progressive motility (pre-freeze and post-thaw) and the mean of fertilization rates of MBCD-GSH IVF. Ypre = pre-freeze progressive motility; Ypost = post-thaw progressive motility; X = IVF rate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940049&req=5

Figure 2: Linear regression between the mean of sperm progressive motility (pre-freeze and post-thaw) and the mean of fertilization rates of MBCD-GSH IVF. Ypre = pre-freeze progressive motility; Ypost = post-thaw progressive motility; X = IVF rate.
Mentions: Sperm samples of different mouse lines had different sperm motility. To determine the correlation between sperm motility and their fertilization abilities, MBCD-GSH IVF procedures were performed using cryopreserved sperm samples from 46 knockout mouse lines on a C57BL/6N genetic background. Although there was a large degree of variation in the IVF rate of thawed mutant sperm samples for each progressive motility (pre-freeze or post-thaw), there was a significant linear correlation between the mean of the IVF rate and the mean of the progressive motility after the fertilization rates of the 46 IVF procedures (1 IVF procedure per mouse line using sperm of a heterozygous male) were sorted and separated into 5 categories: 5–19%, 20–29%, 30–39%, 40–59%, and 60–80% (Figure 1). The R2 were 0.9623 and 0.9993 for pre-freeze and post-thaw progressive motility, respectively (Figure 2). The higher the mean of the progressive motility (especially the post-thaw progressive motility), the higher the mean of the IVF rate. IVF rates were usually less than 30% when post-thaw progressive sperm motility was less than 10%. The linear equations for the two variables (X, IVF rate; Y, pre-freeze or post-thaw progressive motility) containing slopes and Y-intercepts when X=0 are shown in Figure 2.

Bottom Line: Different protocols incorporating methyl-β-cyclodextrin (MBCD) and reduced glutathione (GSH) have been reported to improve IVF recovery of cryopreserved mouse sperm on a C57BL/6 (J and N) genetic background.Therefore, in the present study we correlated sperm motility with fertilization rate and compared the efficiency of different IVF methods using various sperm samples so as to establish general guidelines for mouse sperm cryorecovery by IVF.Moreover, the presence of a large number of immotile sperm in the sperm-oocyte co-incubation drop was found to reduce IVF success which could be partially reversed by supplementation using monothioglycerol (MTG) during centrifugation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Mouse Biology Program, University of California, Davis, CA 95618, United States.

ABSTRACT

Different protocols incorporating methyl-β-cyclodextrin (MBCD) and reduced glutathione (GSH) have been reported to improve IVF recovery of cryopreserved mouse sperm on a C57BL/6 (J and N) genetic background. However, it is not clear which IVF protocol is most appropriate when using the various methods to cryorecover sperm with different sperm quality and sample volumes. Therefore, in the present study we correlated sperm motility with fertilization rate and compared the efficiency of different IVF methods using various sperm samples so as to establish general guidelines for mouse sperm cryorecovery by IVF. High linear correlation between sperm fertilization rate and progressive motility was found, R(2) was 0.9623 and 0.9993 for pre-freezing and post-thaw progressive motility, respectively. High amounts of cryoprotective agent (CPA) were observed to impair both sperm capacitation and fertilization. Moreover, the presence of a large number of immotile sperm in the sperm-oocyte co-incubation drop was found to reduce IVF success which could be partially reversed by supplementation using monothioglycerol (MTG) during centrifugation. It was concluded that the efficiency of IVF using cryorecovered mouse sperm in media containing MBCD and GSH can be predicted from sperm progressive motility. High concentrations of CPA and immotile sperm should be mitigated prior to IVF. The optimum IVF method should be selected based on sperm sample volume and sperm parameters.

No MeSH data available.


Related in: MedlinePlus