早孕因子阳性血清添加到牛体外受精培养基中的补充效果。

Supplementation effect of early pregnancy factor-positive serum into bovine in vitro fertilization culture medium.

作者信息

Ito K, Takahashi M, Kawahata K, Goto T, Takahashi J, Yasuda Y

机构信息

Department of Bioscience and Technology, Faculty of Agriculture, Iwate University, Japan.

出版信息

Am J Reprod Immunol. 1998 Jun;39(6):356-61. doi: 10.1111/j.1600-0897.1998.tb00369.x.

DOI:10.1111/j.1600-0897.1998.tb00369.x

PMID:9645265

Abstract

PROBLEM

Early pregnancy factor (EPF) has been detected in pregnant bovine serum, and its activity appeared from 24 to 48 hr after insemination. However, in bovine in vitro fertilization (IVF), an EPF-like substance(s) had been detected in the culture medium of fertilized ovum. Therefore, we think that EPF and EPF-like substance(s) are very important materials for the development of the embryo. In this study, we examined the development of the embryo when fertilized bovine ova were cultured with IVF culture medium supplemented with EPF-positive or -negative serum.

METHOD OF STUDY

EPF activity of each serum (fetal calf serum [FCS], calf serum [CS], estrus bovine serum, and pregnant bovine serum) was assessed by the bovine-rosette inhibition test. The sera were supplemented in TCM-199 culture medium, and IVF bovine ova were cultured with the media for 6 or 7 days, respectively. The culture media of each group were evaluated for EPF activity by the bovine-rosette inhibition test 48 hr after IVF. The cleavage rate of each group was calculated at 48 hr, and 6 or 7 days after IVF. The culture medium of cumulus cells was also tested for EPF activity.

RESULTS

Only the pregnant bovine sera were EPF positive. All the culture media 48 hr after IVF became EPF positive. Additionally, the culture medium of cumulus cells did not have EPF activity. There was no significant difference in the cleavage rate of the EPF-positive and -negative sera 48 hr after IVF. However, the cleavage rate of EPF-positive sera tended to be higher than the negative sera. In contrast, the blastocyst development rates of EPF-positive sera were significantly higher than those of the negative sera 6 to 7 days after IVF (P < 0.05).

CONCLUSIONS

The data suggest that an EPF-like substance(s) may be secreted from the in vitro fertilized bovine ovum but not from the cumulus cell, and that the EPF in the pregnant serum may accelerate the development of the bovine embryo in IVF.

摘要

问题

已在怀孕母牛血清中检测到早期妊娠因子（EPF），其活性在授精后24至48小时出现。然而，在牛体外受精（IVF）中，已在受精卵的培养基中检测到一种类似EPF的物质。因此，我们认为EPF和类似EPF的物质是胚胎发育的非常重要的物质。在本研究中，我们检查了用补充有EPF阳性或阴性血清的IVF培养基培养受精牛卵时胚胎的发育情况。

研究方法

通过牛红细胞凝集抑制试验评估每种血清（胎牛血清 [FCS]、小牛血清 [CS]、发情期母牛血清和怀孕母牛血清）的EPF活性。将血清添加到TCM-199培养基中，IVF牛卵分别用这些培养基培养6或7天。在IVF后48小时，通过牛红细胞凝集抑制试验评估每组培养基的EPF活性。在IVF后48小时、6或7天计算每组的卵裂率。还检测了卵丘细胞培养基的EPF活性。

结果

只有怀孕母牛血清的EPF呈阳性。IVF后48小时所有培养基的EPF均呈阳性。此外，卵丘细胞培养基没有EPF活性。IVF后48小时，EPF阳性和阴性血清的卵裂率没有显著差异。然而，EPF阳性血清的卵裂率往往高于阴性血清。相比之下，IVF后6至7天，EPF阳性血清的囊胚发育率显著高于阴性血清（P < 0.05）。