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体内受精与体外受精的家猫胚胎的发育能力

Developmental competence of domestic cat embryos fertilized in vivo versus in vitro.

作者信息

Roth T L, Swanson W F, Wildt D E

机构信息

National Zoological Park, Smithsonian Institution, Washington, District of Columbia 20008.

出版信息

Biol Reprod. 1994 Sep;51(3):441-51. doi: 10.1095/biolreprod51.3.441.

Abstract

Development of in vitro-fertilized (IVF) cat embryos was compared to that of naturally produced cat embryos in vivo and in vitro. To obtain in vivo-fertilized embryos, queens were mated three times daily on the second and third days of natural estrus and ovariohysterectomized at 64, 76, 100, 124, or 148 h after the first copulation. Embryos were flushed from the reproductive tract, evaluated for developmental stage, and cultured. For IVF, oocytes from gonadotropin-stimulated queens were inseminated with electroejaculated cat sperm in Ham's F-10 and evaluated for fertilization (cleavage to > or = 2 cells) at 30 h. In vitro development of embryos fertilized in vivo (n = 109) and in vitro (n = 46) was evaluated every 24 h for up to 10 days. High-quality embryos recovered at 64, 76, 100, 124, and 148 h after the first copulation were typically 1 to 2 cells (13 of 20), 5 to 8 cells (18 of 28), 9 to 16 cells (14 of 24), morulae (15 of 21), and compact morulae (11 of 18), respectively, suggesting blastomere cleavage once per day in vivo after the first three rapid cell divisions. A similar developmental rate to the morula stage (p > or = 0.05) was achieved in vitro by embryos derived from both in vitro and in vivo fertilization. Additionally, the proportion (p > or = 0.05) of in vivo-generated embryos (2 to 16 cells) that developed to morulae (64 of 83; 77.1%) was similar to that of IVF embryos (28 of 46; 60.9%). However, none of the IVF embryos (0/46), but 70.6% (77 of 109) of the in vivo-produced embryos, achieved blastocyst formation in culture (p < or = 0.05). Furthermore, 66.2% (51 of 77) of these blastocysts exhibited zona hatching. Incidence of morula and blastocyst formation in the in vivo group was influenced by stage of the embryo at collection. Embryos that were at the 9- to 16-cell stage at recovery were more likely (p < or = 0.05) to achieve morula or blastocyst status and emerge from the zona pellucida than younger-stage counterparts. In summary, the in vivo and in vitro growth rate of cat embryos produced after natural mating was comparable to that of embryos fertilized and cultured in vitro. However, developmental ability to the blastocyst stage was superior for embryos produced in vivo after natural mating.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

将体外受精(IVF)的猫胚胎的发育情况与体内和体外自然产生的猫胚胎进行了比较。为了获得体内受精的胚胎,在自然发情的第二天和第三天,每天让母猫与公猫交配三次,并在第一次交配后64、76、100、124或148小时进行卵巢子宫切除术。从生殖道中冲洗出胚胎,评估其发育阶段并进行培养。对于IVF,用促性腺激素刺激的母猫的卵母细胞在Ham's F - 10中与电刺激采集的猫精子进行受精,并在30小时时评估受精情况(卵裂至≥2个细胞)。对体内受精(n = 109)和体外受精(n = 46)的胚胎进行长达10天的体外发育评估,每24小时评估一次。在第一次交配后64、76、100、124和148小时回收的高质量胚胎通常分别为1至2细胞期(20个中的13个)、5至8细胞期(28个中的18个)、9至16细胞期(24个中的14个)、桑葚胚期(21个中的15个)和致密桑葚胚期(18个中的11个),这表明在前三次快速细胞分裂后,体内每天进行一次卵裂球分裂。来自体外受精和体内受精的胚胎在体外发育到桑葚胚期的速率相似(p≥0.05)。此外,体内产生的胚胎(2至16细胞期)发育到桑葚胚期的比例(83个中的64个;77.1%)与IVF胚胎(46个中的28个;60.9%)相似(p≥0.05)。然而,在培养过程中,没有IVF胚胎(0/46)形成囊胚,但70.6%(109个中的77个)的体内产生的胚胎形成了囊胚(p≤0.05)。此外,这些囊胚中有66.2%(77个中的51个)出现透明带孵化。体内组中桑葚胚和囊胚形成的发生率受采集时胚胎阶段的影响。回收时处于9至16细胞期的胚胎比早期阶段的胚胎更有可能(p≤0.05)发育到桑葚胚或囊胚阶段并从透明带中孵出。总之,自然交配后产生的猫胚胎在体内和体外的生长速率与体外受精和培养的胚胎相当。然而,自然交配后体内产生的胚胎发育到囊胚阶段的能力更强。(摘要截断于400字)

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