Shirazi Abolfazl, Motaghi Ehsan
Embryology & Andrology Department of Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran ; Department of Gametes and Cloning, Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.
J Reprod Infertil. 2013 Jan;14(1):8-16.
The cells of mammalian female reproductive tract have been widely used for in vitro fertilization (IVF). This study was designed to study the effects of oviductal epithelial cells (OECs) and their conditioned medium during IVF on subsequent embryonic development and the relative abundance of zygote arrest 1 (Zar1) transcript in ovine zygotes.
The in vitro matured ovine oocytes were randomly fertilized in the following culture conditions: I) SOFaaBSA+20% sheep serum (control), II) SOFaa BSA+20% sheep serum (50 µl) in the presence of OECs, III) SOFaaBSA+20% sheep serum (100 µl) in the presence of OECs, and IV) OECs conditioned medium (CM). Sigma Stat (Version 2.0) software and one-way ANOVA were considered for statistical analysis. A p<0.05 was considered statistically significant.
The cleavage, blastocyst, and hatched blastocyst rates in OECs and CM groups were significantly lower than the control group (p<0.01). In co-cultured groups, the application of two different volumes of IVF medium showed no difference in embryonic developmental indices. The Zar1 gene expression in zygotes produced in the presence of OECs was significantly higher than those produced in the control and CM groups (p<0.05).
Neither the presence of oviductal epithelial cells nor their conditioned medium could improve the developmental potential of ovine embryos during IVF. Moreover, no relationship was observed between the relative abundance of Zar1 transcript in zygotes produced in different conditions and the corresponding subsequent embryonic development.
哺乳动物雌性生殖道细胞已广泛用于体外受精(IVF)。本研究旨在探讨体外受精过程中输卵管上皮细胞(OECs)及其条件培养基对绵羊受精卵后续胚胎发育以及合子阻滞1(Zar1)转录本相对丰度的影响。
将体外成熟的绵羊卵母细胞在以下培养条件下随机受精:I)SOFaaBSA + 20%绵羊血清(对照组),II)在存在OECs的情况下加入SOFaa BSA + 20%绵羊血清(50 μl),III)在存在OECs的情况下加入SOFaaBSA + 20%绵羊血清(100 μl),IV)OECs条件培养基(CM)。采用Sigma Stat(版本2.0)软件和单因素方差分析进行统计分析。p < 0.05被认为具有统计学意义。
OECs组和CM组的卵裂率、囊胚率和孵化囊胚率均显著低于对照组(p < 0.01)。在共培养组中,两种不同体积的IVF培养基的应用在胚胎发育指标上没有差异。在存在OECs的情况下产生的受精卵中Zar1基因表达显著高于对照组和CM组(p < 0.05)。
输卵管上皮细胞及其条件培养基的存在均不能提高体外受精过程中绵羊胚胎的发育潜能。此外,在不同条件下产生的受精卵中Zar1转录本的相对丰度与相应的后续胚胎发育之间未观察到相关性。