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白蛋白和柠檬酸钠对体外生产牛胚胎发育的差异需求。

The differential requirement of albumin and sodium citrate on the development of in vitro produced bovine embryos.

作者信息

Sung Li-Ying, Du Fuliang, Xu Jie, Chang Wancun, Nedambale Tshimangadzo Lucky, Zhang Jiaxin, Jiang Shie, Tian X Cindy, Yang Xiangzhong

机构信息

Department of Animal Science and Connecticut Center for Regenerative Biology, University of Connecticut, Storrs, CT 06269, USA.

出版信息

Reprod Nutr Dev. 2004 Nov-Dec;44(6):551-64. doi: 10.1051/rnd:2004061.

Abstract

In vitro culture for bovine embryos is largely not optimal. Our study was to determine the components necessary for early embryo development. In experiment 1, IVF embryos were cultured for two days in CR1aa medium containing sodium citrate and BSA from two sources (Sigma vs. ICPbio), subsequently for additional five days with cumulus monolayer in 10% FBS CR1aa. We found that supplementation with both Sigma-BSA and sodium citrate significantly increased total blastocyst (BL) development compared with the ICPbio-BSA groups (37% vs. 19-21%), and enhanced the total number of high quality (C1 BL, IETS standard) blastocysts (26% vs. 11-17%) (P < 0.05). In experiment 2 with serum free and/or somatic free culture, we found that CR1aa culture can support a comparable embryo development with a supplement of Sigma BSA. The addition of sodium citrate did not increase blastocyst development in either the Sigma-BSA or the ICPbio-BSA groups. An inferior blastocyst development occurring in ICPbio-BSA culture (1-3%) could be rescued by culture in CRlaa supplemented with 10% FBS (29%), more importantly, by culture in CR1aa with a replacement of Sigma BSA (24%) (P <0.05). C1 blastocysts rescued by FBS and Sigma BSA in ICPbio-BSA culture possessed indistinguishable morphology to embryos developed in a Sigma-BSA, FBS and somatic co-culture system, showing similar cell number/blastocyst (129-180, P > 0.05). Our study found a beneficial effect of sodium citrate and BSA on the in vitro development of bovine IVF embryos during co-culture. We also determined that differential embryotrophic factor(s) contained in BSA and serum, probably not sodium citrate, is necessary for promoting competent morula and blastocyst development in cattle.

摘要

牛胚胎的体外培养在很大程度上并不理想。我们的研究旨在确定早期胚胎发育所需的成分。在实验1中,体外受精胚胎在含有柠檬酸钠和两种来源(Sigma与ICPbio)牛血清白蛋白(BSA)的CR1aa培养基中培养两天,随后在含有10%胎牛血清(FBS)的CR1aa培养基中与卵丘单层共同培养另外五天。我们发现,与使用ICPbio-BSA的组相比,添加Sigma-BSA和柠檬酸钠显著提高了囊胚的总体发育率(37%对19%-21%),并增加了高质量(C1囊胚,符合IETS标准)囊胚的总数(26%对11%-17%)(P<0.05)。在实验2的无血清和/或无体细胞培养中,我们发现CR1aa培养基在添加Sigma BSA的情况下能够支持相当的胚胎发育。添加柠檬酸钠在Sigma-BSA组或ICPbio-BSA组中均未增加囊胚发育。在ICPbio-BSA培养基中出现的较差囊胚发育率(1%-3%)可通过在添加10%FBS的CRlaa培养基中培养(29%)得到挽救,更重要的是,通过在CR1aa培养基中用Sigma BSA替代(24%)得到挽救(P<0.05)。在ICPbio-BSA培养基中由FBS和Sigma BSA挽救的C1囊胚与在Sigma-BSA、FBS和体细胞共培养系统中发育的胚胎具有难以区分的形态,显示出相似的每个囊胚细胞数(129-180,P>0.05)。我们的研究发现柠檬酸钠和BSA在共培养期间对牛体外受精胚胎的体外发育具有有益作用。我们还确定,BSA和血清中含有的不同胚胎营养因子(可能不是柠檬酸钠)对于促进牛的有能力桑椹胚和囊胚发育是必要的。

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