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Effect of Culture Conditions on Viability of Mouse and Rat Embryos Developed in Vitro

机译:培养条件对体外发育的小鼠和大鼠胚胎存活力的影响

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摘要

Currently in vitro culture of mouse preimplantation embryos has become a very important technique to investigate different mechanisms of early embryogenesis. However, there is a big difference in the preimplantation development between mammalian species. Despite close relatedness to mice, in vitro cultivation of rat preimplantation embryos is still delicate and needs further investigation and optimizations. In this study we have compared the in vitro developmental potential of mouse and rat embryos cultured at different culture conditions in parallel experiments. Interestingly, mouse zygotes developed in vitro until blastocyst stage even in inadequate medium without any phosphates and with low osmolarity which was formulated especially for cultivation of rat embryos. Rat parthenotes and zygotes developed in M16 medium formulated for mouse embryos only till 2-cell stage and further development is blocked completely at this stage. Moreover, developmental ability of rat embryos in vitro was significantly lower in comparison with mouse even in special rat mR1ECM medium. Mouse and rat embryos at 2-cell stage obtained in vivo developed until blastocyst stages significantly more efficiently compared to zygotes. Culture of mouse zygotes in glass capillaries resulted in a significantly higher rate of morula and blastocyst development compared with dishes. The Well-of-the-Well system resulted in a significant improvement when compared with dishes for the culture of rat zygotes only until morula stage. Reduced oxygen tension increased the developmental rate of rat but not mouse zygotes until blastocyst stage. This study demonstrates that development of early preimplantation embryos is altered by different culture conditions and show strong differences even between two related species such as mice and rats. Therefore, for understanding the fundamental mechanisms of early mammalian development it is very important to use embryos of various species.
机译:目前,小鼠植入前胚胎的体外培养已成为研究早期胚胎发生的不同机制的非常重要的技术。但是,哺乳动物之间的植入前发育有很大差异。尽管与小鼠密切相关,但大鼠体外植入前胚胎的体外培养仍然微妙,需要进一步研究和优化。在这项研究中,我们在平行实验中比较了在不同培养条件下培养的小鼠和大鼠胚胎的体外发育潜力。有趣的是,小鼠受精卵在体外发育直至囊胚期,甚至在没有任何磷酸盐且具有低渗透压的不适当培养基中也是如此,这是专门为培养大鼠胚胎而配制的。在为小鼠胚胎配制的M16培养基中发育的大鼠单性生殖和合子仅持续到2细胞阶段,并且在该阶段完全阻止了进一步的发育。此外,即使在特殊的大鼠mR1ECM培养基中,大鼠胚胎的体外发育能力也明显低于小鼠。与合子相比,在体内获得的2细胞阶段的小鼠和大鼠胚胎发育到胚泡阶段显着更有效。与皿相比,在玻璃毛细管中培养小鼠受精卵导致桑mor和胚泡发育的比率显着更高。与仅培养桑直到大鼠受精卵的培养皿相比,Well-of-Well系统产生了显着改善。降低的氧张力增加了大鼠受精卵的发育速度,但直到囊胚期才增加。这项研究表明,早期的植入前胚胎的发育会因不同的培养条件而改变,甚至在两个相关物种(如小鼠和大鼠)之间也显示出很大的差异。因此,为了理解哺乳动物早期发育的基本机制,使用各种物种的胚胎非常重要。

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