Development of porcine embryos from one- and two-cell stages to blastocysts in culture medium supplemented with porcine oviductal fluid.
Development of porcine embryos from one- and two-cell stages to blastocysts in culture medium supplemented with porcine oviductal fluid.
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在补充有猪输卵管液的培养基中猪胚胎从一细胞和二细胞阶段发育成囊胚。
DOI：
10.1095/biolreprod41.6.1076
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发表时间：
1989
影响因子：
3.6
通讯作者：
Johnson,BH
中科院分区：
文献类型：
--
作者：
Archibong,AE;Petters,RM;Johnson,BH
从发情周期第1天（发情周期第0天=检测到发情的当天）开始，从5头母猪中长期采集输卵管液（OVF），并用于胚胎培养（仅第3天OVF）。进行了两个实验以研究1-细胞和2-细胞猪胚胎在改良的Kreb’s Ringer碳酸氢盐培养基（培养基，CM）、早期黄体期OVF或补充有OVF的CM（CM-OVF，CM中25%OVF v/v）中的体外发育，并转移或不转移到新鲜CM。在实验1中，在检测到发情后约44 h，从母猪（n = 7）中收获1-细胞和2-细胞胚胎。在实验2中，在注射人绒毛膜促性腺激素后约50 h，从5头用烯丙孕素和促性腺激素处理的母猪中收集1-细胞胚胎。OVF体积（ml）在采集的4天内逐渐下降（发情开始后24 h，8.44 ± 0.28; 48 h，6.88 ± 1.78; 72 h，4.96 ± 0.35; 96 h，4.64 ± 0.25; p <0.01）。在两个实验中，在OVF中培养的胚胎中发育至囊胚期的比例最低，在CM-OVF中培养的胚胎中发育至囊胚期的比例最高（实验1：CM，27.3; OVF，10; CM-OVF，63.6;实验2：CM，26.7; OVF，0; CM-OVF，82.4;囊胚形成%）。当胚胎暴露于OVF和CM-OVF 48 h（1-细胞胚胎）或24 h（2-细胞胚胎）转移到新鲜CM时，在实验I中发育到囊胚阶段得到改善（CM到CM，27.3; OVF到CM，58.3; CM-OVF到CM，100;囊胚形成%）。在实验2中，只有从OVF转移到新鲜CM的胚胎显示出囊胚形成百分比的提高（CM到CM，29.3; OVF到CM，35.3; CM-OVF到CM，77.8）。早期黄体期猪OVF本身在体外抑制1-细胞和2-细胞猪胚胎的发育。
Oviductal fluid (OVF) was harvested chronically from 5 sows beginning on Day 1 of the estrous cycle (Day 0 of estrous cycle = day of detected estrus) and used for embryo culture (Day 3 OVF only). Two experiments were conducted to investigate in vitro development of 1-cell and 2-cell porcine embryos in a modified Kreb's Ringer bicarbonate medium (culture medium, CM), early luteal phase OVF or CM supplemented with OVF (CM-OVF, 25% OVF v/v in CM) with or without transfer to fresh CM. In Experiment 1, 1-cell and 2-cell embryos were harvested from sows (n = 7) approximately 44 h after detected estrus. In Experiment 2, 1-cell embryos were collected from 5 sows treated with altrenogest and gonadotropins, approximately 50 h after injection of human chorionic gonadotropin. The volume of OVF (ml) declined progressively throughout the 4 days of collection (24 h, 8.44 ± 0.28; 48 h, 6.88 ± 1.78; 72 h, 4.96 ± 0.35; 96 h, 4.64 ± 0.25 after onset of estrus; p < .01). In both experiments, development to blastocyst stage was lowest among embryos cultured in OVF and highest among those cultured in CM-OVF (Experiment 1: CM, 27.3; OVF, 10; CM-OVF, 63.6; Experiment 2: CM, 26.7; OVF, 0; CM-OVF, 82.4; % blastocyst formation). When embryos exposed to OVF and CM-OVF for 48 h (1-cell embryos) or 24 h (2-cell embryos) were transferred to fresh CM, development to blastocyst stage improved (CM to CM, 27.3; OVF to CM, 58.3; CM-OVF to CM, 100; % blastocyst formation) in Experiment I. In Experiment 2, only embryos transferred from OVF to fresh CM showed improvement in percentage blastocyst formation (CM to CM, 29.3; OVF to CM, 35.3; CM-OVF to CM, 77.8).This study indicates that addition of OVF to culture medium results in improved development in vitro of porcine embryos from 1-cell to blastocyst. Early luteal phase porcine OVF by itself inhibits the development of 1-cell and 2-cell porcine embryos in vitro.
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