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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.


Comparisons of cryorecovery IVF rates of different IVF methods using wildtype C57BL/6N sperm. Each IVF procedure was repeated 3 times. Compared with the other 3 IVF methods, the IVF rate of the “IVF in MBCD” method was significantly lower (P<0.05).
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Figure 3: Comparisons of cryorecovery IVF rates of different IVF methods using wildtype C57BL/6N sperm. Each IVF procedure was repeated 3 times. Compared with the other 3 IVF methods, the IVF rate of the “IVF in MBCD” method was significantly lower (P<0.05).

Mentions: The post-thaw sperm progressive motility of many cryopreserved mutant sperm samples on C57BL/6 backgrounds was less than 10%. To determine which IVF method should be used for a given sperm sample with a post-thaw progressive motility in the range of 5–10% and a sperm concentration of 30–50 × 106/mL, we compared the IVF rates of 4 different IVF methods using 10 µL of C57BL/6N wildtype sperm with post-thaw progressive motility 5–8% from the same sperm pools (mixture of sperm from 3–5 males) for each IVF procedure. As shown in Figure 3, there was no significant difference in IVF rate among 3 of the IVF methods (MBCD-GSH IVF, Rescue MBCD-GSH IVF, and Rescue IVF in MBCD), but the IVF rate of the “IVF in MBCD” method was significantly lower than that of any of the other 3 IVF methods (P<0.05).


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)

Comparisons of cryorecovery IVF rates of different IVF methods using wildtype C57BL/6N sperm. Each IVF procedure was repeated 3 times. Compared with the other 3 IVF methods, the IVF rate of the “IVF in MBCD” method was significantly lower (P<0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Comparisons of cryorecovery IVF rates of different IVF methods using wildtype C57BL/6N sperm. Each IVF procedure was repeated 3 times. Compared with the other 3 IVF methods, the IVF rate of the “IVF in MBCD” method was significantly lower (P<0.05).
Mentions: The post-thaw sperm progressive motility of many cryopreserved mutant sperm samples on C57BL/6 backgrounds was less than 10%. To determine which IVF method should be used for a given sperm sample with a post-thaw progressive motility in the range of 5–10% and a sperm concentration of 30–50 × 106/mL, we compared the IVF rates of 4 different IVF methods using 10 µL of C57BL/6N wildtype sperm with post-thaw progressive motility 5–8% from the same sperm pools (mixture of sperm from 3–5 males) for each IVF procedure. As shown in Figure 3, there was no significant difference in IVF rate among 3 of the IVF methods (MBCD-GSH IVF, Rescue MBCD-GSH IVF, and Rescue IVF in MBCD), but the IVF rate of the “IVF in MBCD” method was significantly lower than that of any of the other 3 IVF methods (P<0.05).

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.