两种抗冷冻剂对不同品系小鼠精子冷冻的影响

Effect of Two Cryoprotectants on Cryopreservation of Different Strains Mouse Sperms

目的建立一套比较理想的小鼠精子冷冻复苏的方法。方法采用R18S3和FERTIUPTM-CPA两种保护剂,对DBA/2、C57BL/6J、KM和B6.129S7-Ldlrtm1Her/J四个品系的小鼠精子进行冷冻,冷冻精子用三种方法复苏,以体外受精率来评价精子冷冻的效果。结果以R18S3作为冷冻保护剂,DBA/2(73.3%,88.4%,55.6%)和KM(64.9%,60.2%,39.6%)品系小鼠冷冻精子复苏后体外受精率差异显著(P<0.05),C57BL/6J(3.0%,10.3%,3.7%)和B6.129S7-Ldlrtm1Her/J(0%,5.0%,0%)结果差异不显著(P>0.05);以FERTIUPTM-CPA作为冷冻保护剂,DBA/2(33.6%,14.1%,91.6%)、C57BL/6J(8.4%,21.0%,4.9%)和B6.129S7-Ldlrtm1Her/J(8.2%,10.0%,28.9%)品系小鼠冷冻精子复苏后体外受精率差异显著(P<0.05),而KM(48.1%,48.0%,48.1%)小鼠冷冻精子复苏后体外受精率差异不显著(P>0.05)。结论对于DBA/2和KM品系小鼠来说,用R18S3或FERTIUPTM-CPA冷冻精子,选择一种恰当的复苏方法,均可以得到较理想的体外受精率,而C57BL/6J和B6.129S7-Ldlrtm1Her/J品系小鼠无论采用哪种冷冻保护剂,选择何种方法复苏精子,得到的体外受精率都较低。

Objective To establish an optimal method of frozen-thawing mouse sperms. Methods Rl8 Ss and FERTIUpTM-cPA were used as cryopreservation protection solutions to freeze mouse sperms of DBA/2, C57BL/6J, KM and B6. 129S7-Ldl^rtalHer/J four strains. Three thawing methods were used and the rate of in vitro fertilization （IVF） was used to assess the result of cryopreservation of sperms. Results When R18 S3 was used to freeze sperms, there was a significant difference between the rates of IVF in DBA/2 （73.3%, 88.4%, 55.6% ）and KM（64.9%, 60.2%, 39.6% ）among three thawing methods （ P 〈0.05）, but not significant difference between C57BL/6J （3.0%, 10.3%, 10.3%） and B6. 129S7-Ldl^raiHer/J （0%, 5. 0% , 0% ）（ P 〉 0.05）. When FERTIUPTM-CPA was used for cryopreservation of sperms, there was a significant difference of the ratios of IVF in DBA/2 （33.6%, 14.1% ,91.6% ）, C57BL/6J （ 8.4% ,21.0% ,4.9% ） and B6. 129S7-Ldlr^tmlHer/J （8.2% , 10.0%, 28.9%）（P〈0.05）, but not with that of KM （48.1%, 48.0%, 48.1%） mice （P〉0.05）. Conclusion For DBA/2 and KM mouse sperms, frozen with either RisSs or FERTIUP^TM-cPA and thawed with a suitable method, both can get an ideal rate of IVF. However, no matter which cryopreservation protective solution and thawing method are used, the rate of IVF of C57BL/6J and B6.129S7-Ldl^rtml Her/J thawed sperms is always relatively decreased.