Hoelker M, Rings F, Lund Q, Phatsara C, Schellander K, Tesfaye D
Institute of Animal Science; Animal Breeding and Husbandry Group, University of Bonn, Bonn, Germany.
Reprod Domest Anim. 2010 Oct;45(5):e138-45. doi: 10.1111/j.1439-0531.2009.01535.x.
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 μm reached significantly higher blastocyst rates (32.5 vs 21.1% vs 20.3%) compared to embryos cultured in WOWs of 300 μm diameter or plain cultured controls. Embryos cultured in WOWs with 700 μm depth reached significant higher developmental rates compared with embryos cultured in WOWs of 300 μm 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 μl was identified to be optimal when the embryo per WOW volume was 1 : 0.27 μl 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.
为克服单胚胎培养导致的发育问题，已开发出“阱中阱”（WOW）培养系统。在本研究中，我们旨在研究胚胎密度在微环境和宏观环境方面对发育率和胚胎质量的影响，以更深入了解具有发育重要性的机制。WOW的直径和深度显著影响发育率（p < 0.05）。与在直径300μm的WOW中培养的胚胎或常规培养的对照组相比，直径500μm的WOW中胚胎的囊胚率显著更高（32.5% 对21.1% 对20.3%）。与在300μm深度的WOW中培养的胚胎及对照胚胎相比，在700μm深度的WOW中培养的胚胎发育率显著更高（30.6% 对22.6% 对20.3%）。每个WOW体积中的胚胎数与发育率的相关性高于WOW直径或深度与发育率的相关性（r(2) = 0.92，p = 0.0004）。然而，每个WOW体积中的胚胎数并不影响细胞计数差异。当每个WOW体积中的胚胎数为1 : 0.27μl时，确定每个培养皿体积中胚胎数为1 : 30μl为最佳，可将发育率提高至批量胚胎生产水平。本研究结果能够在整个培养期间追踪每个胚胎，同时保持高发育率和正常的有丝分裂动态，将为人类辅助生殖中的胚胎单培养、具有高经济价值的哺乳动物胚胎以及科学研究带来益处。
