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Ham's F10与二氧化碳或Hepes缓冲液在5℃下储存马胚胎24小时的比较。

Comparison of Ham's F10 with CO2 or Hepes buffer for storage of equine embryos at 5 C for 24 H.

作者信息

Carnevale E M, Squires E L, McKinnon A O

机构信息

Anim. Reprod. Lab., Colorado State University, Fort Collins 80523.

出版信息

J Anim Sci. 1987 Dec;65(6):1775-81. doi: 10.2527/jas1987.6561775x.

Abstract

Forty equine embryos collected 7 d post-ovulation were stored at 5 C for 24 h in one of two culture media (n = 20/group): 1) Ham's F10 + 10% heat-treated fetal calf serum (FCS) buffered by gassing with 5% CO2, 5% O2 and 90% N2 and 2) Ham's F10 + 10% FCS with Hepes buffer (25 mM). Embryos cultured in Ham's F10 + CO2 maintained a better quality score and had a larger average increase in diameter (+34.8 micron) than embryos stored in Hepes buffered Ham's F10 (-10.2 micron). Embryos were transferred surgically into recipient mares that ovulated -3 to +1 d in relation to the donor mare. Twenty embryos cultured in Dulbecco's phosphate buffered saline + 10% FCS and transferred less than 1 h after collection were used as controls. Pregnancy rates were higher (P less than .05) for embryos stored in Ham's F10 + CO2 (70%, 55%) than for embryos stored in Ham's F10 + Hepes (20%, 15%) at 14 and 35 d, respectively. At 14 d, pregnancy rates for control embryos (90%) were similar (P greater than .05) to pregnancy rates for embryos cultured in Ham's F10 + CO2 (70%); however, by 35 d, pregnancy rates were higher (P less than .05) for controls (80%) than for embryos stored in Ham's F10 + CO2 (55%). It was concluded that Ham's F10 + CO2 was superior to Ham's F10 + Hepes for short-term storage of equine embryos at 5 C, and that satisfactory pregnancy rates could be obtained from transfer of embryos stored in Ham's F10 + CO2 at 5 C for 24 h.

摘要

收集40枚排卵后7天的马胚胎,在两种培养基之一中于5℃保存24小时(每组n = 20):1)Ham's F10 + 10% 经热处理的胎牛血清(FCS),通过用5% CO₂、5% O₂和90% N₂通气缓冲;2)Ham's F10 + 10% FCS 并添加25 mM Hepes缓冲液。与保存在添加Hepes缓冲液的Ham's F10中的胚胎(直径平均增加 -10.2微米)相比,在Ham's F10 + CO₂中培养的胚胎保持了更好的质量评分,且直径平均增加更大(+34.8微米)。胚胎通过手术移植到相对于供体母马排卵时间为 -3至 +1天的受体母马体内。20枚在杜氏磷酸盐缓冲盐水 + 10% FCS中培养并在收集后不到1小时进行移植的胚胎用作对照。在14天和35天时,保存在Ham's F10 + CO₂中的胚胎的妊娠率(分别为70%、55%)高于保存在Ham's F10 + Hepes中的胚胎(分别为20%、15%)(P < 0.05)。在14天时,对照胚胎的妊娠率(90%)与在Ham's F10 + CO₂中培养的胚胎的妊娠率(70%)相似(P > 0.05);然而,到35天时,对照胚胎的妊娠率(80%)高于保存在Ham's F10 + CO₂中的胚胎(55%)(P < 0.05)。得出的结论是,在5℃下短期保存马胚胎时,Ham's F10 + CO₂优于Ham's F10 + Hepes,并且从在5℃下于Ham's F10 + CO₂中保存24小时的胚胎移植可获得令人满意的妊娠率。

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