Titterington J L, Robinson J
Department of Applied Biology, University of Hull, UK.
Hum Reprod. 1996 Dec;11(12):2697-702. doi: 10.1093/oxfordjournals.humrep.a019194.
The effects of cryopreservation, in media containing (FS3+) or omitting (FS3) polyvinylpyrrolidone (PVP) in the form of Percoll (PVP-Percoll), on the survival of 2-cell mouse embryos was studied. Survival and zona pellucida disruption post-thaw, growth (assessed by in-vitro culture until the blastocyst stage) and development in vivo (assessed by implantation and living fetus rates and the birth of live progeny) were all investigated. Initial post-thaw survival showed no statistically significant difference (P > 0.05) between FS3+ (91.1 +/- 9.8%) and FS3 (84.5 +/- 6.6%). However, there was a statistically significant (P < 0.05) reduction in the incidence of zona damage when the freezing solution contained PVP-Percoll compared to the control (3.6 +/- 1.0 and 8.7 +/- 0.6% respectively) and a statistically significant (P < 0.05) greater number of embryos developing in vitro to the blastocyst stage (84.8 +/- 7.1 and 72.3 +/- 6.1% respectively). The rates of implantation were not significantly different: 72.2 +/- 7.0% for FS3+ and 51.2 +/- 30.7% for the non-frozen control group. The percentage of live fetuses was also similar between the experimental and control groups: 27.4 +/- 10.6 and 24.3 +/- 11.3% respectively. We conclude that the presence of polymers can protect embryos against cryoinjury and that PVP in the form of PVP-Percoll provides a non-toxic alternative to PVP in its native form, during the cryopreservation of mouse 2-cell embryos.
研究了在含有(FS3+)或不含(FS3)聚乙烯吡咯烷酮(PVP)(以Percoll形式存在,即PVP-Percoll)的培养基中进行冷冻保存对2细胞期小鼠胚胎存活的影响。研究了解冻后的存活率、透明带破裂情况、生长情况(通过体外培养至囊胚期进行评估)以及体内发育情况(通过着床率、活胎率和活后代出生情况进行评估)。解冻后的初始存活率在FS3+(91.1±9.8%)和FS3(84.5±6.6%)之间无统计学显著差异(P>0.05)。然而,与对照组相比,当冷冻溶液含有PVP-Percoll时,透明带损伤发生率有统计学显著降低(P<0.05)(分别为3.6±1.0%和8.7±0.6%),并且体外发育至囊胚期的胚胎数量有统计学显著增加(P<0.05)(分别为84.8±7.1%和72.3±6.1%)。着床率无显著差异:FS3+为72.2±7.0%,非冷冻对照组为51.2±30.7%。实验组和对照组的活胎百分比也相似:分别为27.4±10.6%和24.3±11.3%。我们得出结论,聚合物的存在可以保护胚胎免受冷冻损伤,并且在小鼠2细胞胚胎冷冻保存过程中,PVP-Percoll形式的PVP提供了一种无毒的替代天然形式PVP的选择。