Lim J M, Hansel W
Department of Reproductive Biotechnology, LSU-Pennington Biomedical Research Center, Baton Rouge 70808-4124, USA.
Reprod Fertil Dev. 1996;8(8):1199-205. doi: 10.1071/rd9961199.
Bovine embryos at the 8- or 16-cell stage were cultured singly, or in groups (10-12 embryos), in the presence or absence of bovine oviduct epithelial cells (BOEC) in a defined medium which was used as a basic culture medium. A higher (P < 0.05) proportion of 8-cell embryos (48.3-50.8%) cultured singly developed beyond the 8-cell stage after the addition of platelet-derived growth factor (PDGF)-AB (1 ng mL-1) only, or with PDGF-AB + basic fibroblast growth factor (bFGF; 1 ng ml-1) + transforming growth factor (TGF)-beta 1 beta 2 (1 ng mL-1) than in basic medium alone (30.3%). In contrast, a significantly (P < 0.02) higher percentage (62.6-65.8%) of 16-cell embryos developed to the morula stage after the addition of TGF-beta 1 beta 2 only, or the addition of TGF-beta 1 beta 2 + bFGF + PDGF-AB than in basic medium alone (30.2%). These proportions were not significantly (P > 0.05) different from the proportions obtained when embryos were cultured in groups, but were significantly (P < 0.005) lower than the proportions obtained when embryos were cultured in groups on BOEC monolayers. Arachidonic acid (50 ng mL-1), beta-mercaptoethanol (10 microM) and glutathione (10-1000 microM) stimulated the development of 8-cell embryos in the presence of PDGF and TGF-beta 1 beta 2; blastocyst formation was observed for the first time in 8-cell embryos cultured singly in the presence of these embryotrophic substances (2.2-6.2%).
将处于8细胞或16细胞阶段的牛胚胎单独培养,或分组培养(每组10 - 12个胚胎),在有或无牛输卵管上皮细胞(BOEC)存在的情况下,于一种特定培养基中培养,该培养基用作基础培养基。单独培养的8细胞胚胎,仅添加血小板衍生生长因子(PDGF)-AB(1 ng/mL),或添加PDGF-AB +碱性成纤维细胞生长因子(bFGF;1 ng/mL)+转化生长因子(TGF)-β1β2(1 ng/mL)后,发育至8细胞阶段以上的比例(48.3 - 50.8%)高于仅在基础培养基中培养的比例(30.3%),差异有统计学意义(P < 0.05)。相比之下,16细胞胚胎仅添加TGF-β1β2,或添加TGF-β1β2 + bFGF + PDGF-AB后,发育至桑葚胚阶段的比例(62.6 - 65.8%)显著高于仅在基础培养基中培养的比例(30.2%),差异有统计学意义(P < 0.02)。这些比例与胚胎分组培养时获得的比例相比,差异无统计学意义(P > 0.05),但显著低于在BOEC单层上分组培养胚胎时获得的比例(P < 0.005)。花生四烯酸(50 ng/mL)、β-巯基乙醇(10 μM)和谷胱甘肽(10 - 1000 μM)在PDGF和TGF-β1β2存在的情况下刺激8细胞胚胎的发育;在这些胚胎营养物质存在下单独培养的8细胞胚胎中首次观察到囊胚形成(2.2 - 6.2%)。