Gardner David K, Lane Michelle
Department of Zoology, The University of Melbourne, Melbourne, VIC, Australia.
Methods Mol Biol. 2014;1092:167-82. doi: 10.1007/978-1-60327-292-6_11.
Since the inception of modern embryo culture media over 50 years ago there have been significant developments in culture systems for the mammalian preimplantation embryo. Carbohydrate gradients have been shown to impact embryo physiology and viability, while amino acids have been determined to have specific temporal effects during the preimplantation period. Furthermore, due to the lability of amino acids at 37 °C and the subsequent release of embryo-toxic ammonium into the medium, the medium is renewed every 48 h not only to provide stage-specific nutrients but also to prevent toxicity. Subsequently stage-specific media are commonly employed. To facilitate the preparation of small volumes of media, whose formulations can be readily altered according to experimental design, the use of stock solutions is described, together with systems to facilitate the development of viable embryos.
自50多年前现代胚胎培养基问世以来，哺乳动物植入前胚胎的培养系统有了重大发展。已表明碳水化合物梯度会影响胚胎生理和活力，而氨基酸在植入前期具有特定的时间效应。此外，由于氨基酸在37°C时不稳定，随后会向培养基中释放胚胎毒性铵，因此每48小时更换一次培养基，不仅是为了提供特定阶段的营养物质，也是为了防止毒性。随后通常使用特定阶段的培养基。为便于制备少量培养基，其配方可根据实验设计轻松更改，本文描述了储备溶液的使用方法以及促进胚胎发育的系统。
