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牛输卵管上皮细胞、胎牛血清和牛血清白蛋白对合成输卵管液培养系统中产生的单个牛胚胎基因表达的影响。

Effects of bovine oviduct epithelial cells, fetal calf serum and bovine serum albumin on gene expression in single bovine embryos produced in the synthetic oviduct fluid culture system.

作者信息

Pedersen Mona E, Øzdas Øzen Banu, Farstad Wenche, Tverdal Aage, Olsaker Ingrid

机构信息

Norwegian School of Veterinary Science, 0033 Oslo, Norway.

出版信息

Reprod Fertil Dev. 2005;17(8):751-7. doi: 10.1071/rd05048.

Abstract

In this study the synthetic oviduct fluid (SOF) system with bovine oviduct epithelial cell (BOEC) co-culture is compared with an SOF system with common protein supplements. One thousand six hundred bovine embryos were cultured in SOF media supplemented with BOEC, fetal calf serum (FCS) and bovine serum albumin (BSA). Eight different culture groups were assigned according to the different supplementation factors. Developmental competence and the expression levels of five genes, namely glucose transporter-1 (Glut-1), heat shock protein 70 (HSP), connexin43 (Cx43), (2)-actin (ACTB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), analysed as mRNA by using reverse transcription-polymerase chain reaction, were measured on bovine embryos cultured for 9 days. Gene expression of these in vitro-produced embryos was compared with the gene expression of in vivo-produced embryos. There was no significant difference found in embryo developmental competence between the Day 9 embryos in BOEC co-culture, FCS and BSA supplements in SOF media. However, differences in gene expression were observed. With respect to gene expression in in vivo and in vitro embryos, BOEC co-culture affected the same genes as did supplementation with FCS and BSA. HSP was the only gene that differed significantly between in vitro and in vivo embryos. When the different in vitro groups were compared, a significant difference between the BOEC co-culture and the FCS supplementation groups due to Glut-1 expression was observed.

摘要

在本研究中,将具有牛输卵管上皮细胞(BOEC)共培养的合成输卵管液(SOF)系统与具有普通蛋白质补充剂的SOF系统进行了比较。1600枚牛胚胎在添加了BOEC、胎牛血清(FCS)和牛血清白蛋白(BSA)的SOF培养基中培养。根据不同的补充因子分配了八个不同的培养组。在培养9天的牛胚胎上,通过逆转录-聚合酶链反应分析了五个基因,即葡萄糖转运蛋白-1(Glut-1)、热休克蛋白70(HSP)、连接蛋白43(Cx43)、β-肌动蛋白(ACTB)和甘油醛-3-磷酸脱氢酶(GAPDH)的发育能力和mRNA表达水平。将这些体外生产胚胎的基因表达与体内生产胚胎的基因表达进行了比较。在SOF培养基中,BOEC共培养、FCS和BSA补充剂处理的第9天胚胎之间,胚胎发育能力没有显著差异。然而,观察到了基因表达的差异。关于体内和体外胚胎的基因表达,BOEC共培养影响的基因与FCS和BSA补充剂影响的基因相同。HSP是体外和体内胚胎之间唯一有显著差异的基因。当比较不同的体外组时,观察到由于Glut-1表达,BOEC共培养组和FCS补充剂组之间存在显著差异。

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