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在分离的小鼠输卵管中培养绵羊体外成熟/体外受精合子:基础培养基的作用。

Culture of Ovine IVM/IVF Zygotes in Isolated Mouse Oviduct: Effect of Basal Medium.

作者信息

Farahavar Abbas, Shirazi Abolfazl, Kohram Hamid, Shahneh Ahmad Zareh, Sarvari Ali, Naderi Mohammad Mehdi, Boroujeni Sara Borjian, Zhandi Mehdi

机构信息

Department of Animal Science, Faculty of Agriculture and Natural Resources, University of Tehran, Karaj, Iran.

出版信息

Avicenna J Med Biotechnol. 2013 Apr;5(2):133-7.

Abstract

BACKGROUND

The basal medium that supports Isolated Mouse Oviduct (IMO) is important for supporting embryo development and quality.

METHODS

The culture of ovine IVM/IVF zygotes was done in IMO using SOFaaciBSA and SOFaaBSA as basal medium of IMO and in SOFaaBSA alone as control. For preparation of IMO mature inbred strain C57BL/6 female mice were synchronized and mated with vasectomized males. The females with vaginal plug were sacrificed and the zygotes were transferred in to the isolated oviduct at 20 hpi. The oviducts were cultured with SOFaaciBSA and SOFaaBSA for 6 days. Another group of zygotes were cultured in SOFaaBSA alone as control.

RESULTS

Culture of zygotes in the IMO with SOFaaciBSA and SOFaaBSA, did not significantly affect the development and quality of embryos (p > 0.05). The hatching rate, total and trophectoderm cells number in IMO groups' blastocysts were significantly higher than SOFaaBSA alone. The morphological appearance of IMO blastocysts was superior to SOFaaBSA alone. When the quality of oocytes was poor, IMO could better support ovine embryo development either with SOFaaBSA or SOFaaciBSA than SOFaaBSA alone and there was a significant difference in blastocyst formation at day 6 with SOFaaBSA alone.

CONCLUSION

The culture of ovine IVM/IVF zygotes in IMO using two highly efficient ruminant embryo culture media not only could support development of ovine embryos similar to the level in non IMO culture system (SOFaaBSA alone) but also could improve the quality of resulting embryos. Additionally, IMO could better support the development of ovine embryos derived from poor quality oocytes compared to the SOFaaBSA alone.

摘要

背景

支持分离的小鼠输卵管(IMO)的基础培养基对于支持胚胎发育和质量很重要。

方法

使用SOFaaciBSA和SOFaaBSA作为IMO的基础培养基,在IMO中进行绵羊体外成熟/体外受精(IVM/IVF)合子的培养,并单独使用SOFaaBSA作为对照。为制备IMO,将成熟的近交系C57BL/6雌性小鼠进行同步化处理,并与输精管切除的雄性小鼠交配。处死有阴道栓的雌性小鼠,并在授精后20小时将合子转移到分离的输卵管中。将输卵管用SOFaaciBSA和SOFaaBSA培养6天。另一组合子单独在SOFaaBSA中培养作为对照。

结果

在含有SOFaaciBSA和SOFaaBSA的IMO中培养合子,对胚胎的发育和质量没有显著影响(p>0.05)。IMO组囊胚的孵化率、总细胞数和滋养外胚层细胞数显著高于单独使用SOFaaBSA的组。IMO囊胚的形态外观优于单独使用SOFaaBSA的组。当卵母细胞质量较差时,与单独使用SOFaaBSA相比,IMO使用SOFaaBSA或SOFaaciBSA能更好地支持绵羊胚胎发育,并且在第6天单独使用SOFaaBSA时囊胚形成有显著差异。

结论

在IMO中使用两种高效的反刍动物胚胎培养基培养绵羊IVM/IVF合子,不仅可以支持绵羊胚胎发育至与非IMO培养系统(单独使用SOFaaBSA)相似的水平,还可以提高所得胚胎的质量。此外,与单独使用SOFaaBSA相比,IMO能更好地支持来自质量较差卵母细胞的绵羊胚胎的发育。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a44d/3689557/08bd24b8cff0/AJMB-5-133-g001.jpg

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