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已建立的细胞系及其条件培养基在体外培养期间支持牛胚胎发育。

Established cell lines and their conditioned media support bovine embryo development during in-vitro culture.

作者信息

Myers M W, Broussard J R, Menezo Y, Prough S G, Blackwell J, Godke R A, Thibodeaux J K

机构信息

In Vitro Fertilization Laboratory, Tulsa Center for Fertility and Women's Health, OK 74104.

出版信息

Hum Reprod. 1994 Oct;9(10):1927-31. doi: 10.1093/oxfordjournals.humrep.a138360.

Abstract

These experiments were conducted to evaluate the ability of different somatic-cell monolayers or conditioned medium from somatic cells for supporting bovine embryo development in vitro. In the first experiment, bovine embryos (2- to 4-cells) were allocated randomly to a control (medium 199 with 10% fetal bovine serum and antibiotics) group or co-cultured with bovine oviduct epithelial (BOEC), buffalo rat liver (BRL), Madin Darby bovine kidney (MDBK) or African green monkey kidney (Vero) cells. In the second experiment, bovine embryos (1-cell) were allocated randomly to the following groups: control medium or conditioned medium from BOEC, BRL, MDBK and Vero monolayers. In both experiments, development to the blastocyst stage was assessed after 8 days of incubation at 39 degrees C and 5% CO2. In Experiment 1, coculture improved development to the blastocyst stage compared with control medium alone, and the highest development was observed after co-culture with BOEC. In Experiment 2, conditioned medium enhanced development to morulae and blastocysts compared with the control medium; however, no differences were detected among different cell supports. These results indicate that both co-culture and conditioned medium from different cell monolayers supported development to the blastocyst stage at a higher efficiency than control medium alone.

摘要

进行这些实验是为了评估不同体细胞单层或体细胞条件培养基在体外支持牛胚胎发育的能力。在第一个实验中,将牛胚胎(2至4细胞期)随机分配至对照组(含10%胎牛血清和抗生素的199培养基)或与牛输卵管上皮细胞(BOEC)、水牛大鼠肝细胞(BRL)、马-达二氏牛肾细胞(MDBK)或非洲绿猴肾细胞(Vero)共培养。在第二个实验中,将牛胚胎(1细胞期)随机分配至以下各组:对照培养基或来自BOEC、BRL、MDBK和Vero单层细胞的条件培养基。在两个实验中,于39℃、5%二氧化碳条件下孵育8天后评估胚胎发育至囊胚阶段的情况。在实验1中,与单独使用对照培养基相比,共培养提高了胚胎发育至囊胚阶段的比例,且与BOEC共培养后观察到最高的发育率。在实验2中,与对照培养基相比,条件培养基促进了胚胎发育至桑葚胚和囊胚阶段;然而,在不同细胞支持物之间未检测到差异。这些结果表明,不同细胞单层的共培养和条件培养基支持胚胎发育至囊胚阶段的效率均高于单独使用对照培养基。

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