Effect of Embryo Density on In vitro Developmental Characteristics of Bovine Preimplantative Embryos with Respect to Micro and Macroenvironments
Effect of Embryo Density on In vitro Developmental Characteristics of Bovine Preimplantative Embryos with Respect to Micro and Macroenvironments
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DOI：
10.1111/j.1439-0531.2009.01535.x
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发表时间：
2010-10-01
影响因子：
1.7
通讯作者：
Tesfaye, D.
中科院分区：
文献类型：
--
作者：
Hoelker, M.;Rings, F.;Tesfaye, D.
为了克服单胚胎培养造成的发育问题，人们开发了井中之井（WOW）培养系统。在这项研究中，我们的目的是研究胚胎密度对发育率和胚胎质量的影响，以更深入地了解发育的重要机制。WOW直径和深度显著影响发育率（p < 0.05）。与300 μ m直径的WOW或普通培养对照相比，直径为500 μ m的WOW达到了显著更高的囊胚率（32.5 vs 21.1% vs 20.3%）。与300 μ m深的WOW和对照胚胎相比，在700 μ m深的WOW中培养的胚胎的发育率显著更高（30.6% vs 22.6% vs 20.3%）。胚胎/WOW体积与发育率的相关性高于WOW直径或WOW深度与发育率的相关性（r（2）= 0.92，p = 0.0004）。然而，每WOW体积的胚胎不影响分类细胞计数。当每WOW体积的胚胎为1：0.27 μ l时，每培养皿体积的胚胎为1：30 μ l被确定为最佳的，从而将发育速率提高到大规模胚胎生产的水平。在整个培养期间有机会跟踪每个胚胎，同时保持正常有丝分裂动力学的高发育率，这项工作的结果将为人类辅助生殖中的胚胎单一培养，具有高经济利益的哺乳动物胚胎以及科学目的提供益处。
To overcome developmental problems as a consequence of single embryo culture, the Well of the Well (WOW) culture system has been developed. In this study, we aimed to examine the effect of embryo densities with respect to both microenvironment and macroenvironment on developmental rates and embryo quality to get a deeper insight into developmentally important mechanisms. WOW diameter and depth significantly affected developmental rates (p < 0.05). WOWs with diameter of 500 mu m reached significantly higher blastocyst rates (32.5 vs 21.1% vs 20.3%) compared to embryos cultured in WOWs of 300 mu m diameter or plain cultured controls. Embryos cultured in WOWs with 700 mu m depth reached significant higher developmental rates compared with embryos cultured in WOWs of 300 mu m depth and control embryos (30.6 vs 22.6% vs 20.3%). Correlation of the embryo per WOW volume with developmental rates was higher (r(2) = 0.92, p = 0.0004) than correlation of WOW diameter or WOW depth with developmental rates. However, the embryo per WOW volume did not affect differential cell counts. An embryo per culture dish volume of 1 : 30 mu l was identified to be optimal when the embryo per WOW volume was 1 : 0.27 mu l increasing developmental rates up to the level of mass embryo production. Giving the opportunity to track each embryo over the complete culture period while keeping high developmental rates with normal mitotic dynamics, the results of this work will provide benefit for the single culture of embryos in human assisted reproduction, mammalian embryos with high economic interest as well as for scientific purpose.