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Differences in developmental competence and gene expression profiles between buffalo (Bubalus bubalis) preimplantation embryos cultured in three different embryo culture media

机译:在三种不同的胚胎培养基中培养的水牛(Bubalus bubalis)植入前胚胎的发育能力和基因表达谱的差异

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摘要

The objective of this study was to compare effects of in vitro culture systems on embryonic development and expression patterns of developmentally important genes in preimplantation buffalo embryos. After IVM/IVF presumptive zygotes were cultured in one of three systems: undefined TCM-199, mCR2aa medium supplemented with 10 % FBS and defined PVA–myo-inositol–phosphate–EGF medium. No (P > 0.05) differences at 2-cell, 4-cell and 8-cell to 16- cell stages were observed among the three cultured media used, however, increased (P < 0.05) blastocyst yield, cell number and hatching rate were found in defined medium compared to undefined media. The expression patterns of genes implicated in embryo metabolism (GLUT-1), anti-apoptosis (BCL-2), imprinting (IGF-2R), DNA methylation (DNMT-3A) and maternal recognition of pregnancy (IFNT) were increased (P < 0.05) in hatched blastocysts derived from defined medium compared to undefined media. In conclusion, serum-free, defined medium improved developmental competence of in vitro cultured buffalo embryos. Whether these differences in morphological development and gene expression have long-term effects on buffalo calves born after embryo transfer remains unknown. However, it is possible that early adaptations of the preimplantation embryo to its environment persist during fetal and post-natal development.Electronic supplementary materialThe online version of this article (doi:10.1007/s10616-016-0010-2) contains supplementary material, which is available to authorized users.
机译:这项研究的目的是比较体外培养系统对植入前水牛胚胎中胚胎发育和重要基因的表达模式的影响。将IVM / IVF推定合子在以下三个系统之一中培养后:未定义的TCM-199,补充了10%FBS的mCR2aa培养基和确定的PVA-肌醇-磷酸盐-EGF培养基。在使用的三种培养基中,在2细胞,4细胞和8细胞至16细胞阶段没有观察到(P> 0.05)差异,但是胚泡产量,细胞数量和孵化率增加了(P <0.05)与未定义的媒体相比,在定义的媒体中找到。涉及胚胎代谢(GLUT-1),抗凋亡(BCL-2),印迹(IGF-2R),DNA甲基化(DNMT-3A)和母亲对妊娠的识别(IFNT)的基因的表达模式增加(P小于0.05)来自定义培养基的孵化胚泡与未定义培养基相比。总之,无血清,确定的培养基改善了体外培养的水牛胚胎的发育能力。这些形态发育和基因表达的差异是否对胚胎移植后出生的水牛犊具有长期影响尚不清楚。但是,在胎儿和产后发育过程中,植入前胚胎可能会继续对其环境进行早期适应。电子补充材料本文的在线版本(doi:10.1007 / s10616-016-0010-2)包含补充材料,其中适用于授权用户。

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