Development of preimplantation rabbit embryos after in‐vitro culture and embryo transfer: An electron microscopic study

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Abstract
Compared to in vivo development, in vitro culture of mammalian embryos results in developmental retardation. To study the potential of reversibility of growth retardation we investigated ultrastructurally day 3 rabbit embryos after 1 day in vitro, and cultured embryos that were transferred after culture into recipient rabbits for 1 day. Morphology was compared with ultrastructure of noncultured controls. The noncultured embryos were compacted morulae; the characteristic ultrastructure is described in detail. After 1 day in culture, morulae had developed into early blastocysts. However, unlike the results in vivo, expansion of the blastocysts did not occur and some of the cultured embryos developed trophoblast herniations. In all cultured embryos morphological signs of degeneration were seen with swollen mitochondria, with a dense, granular appearance of the cytoplasm, and with an increase in number of lysosomes. Transfer of cultured blastocysts into uteri of day 3 pseudopregnant recipients resulted in ultrastructurally intact and expanded blastocysts. Transfer into uteri of day 4 pseudopregnant recipients and into uteri of nonpregnant recipients, however, did not yield reversibility of the unphysiological features suffered during the previous time in culture. Although blastocysts were well expanded, distinct signs of injury to the blastomeres were present, proceeding from loss of complete blastomeres to structural changes such as large lamellar structures, dilation of smooth endoplasmic reticulum and Golgi complexes, and clumping of mitochondria. We conclude that developmental retardation during in vitro culture is accompanied by distinct morphological changes. As soon as 24 hr after transfer, these changes can be reversed. This compensation, however, is achieved only if embryos are transferred into recipients that are adapted to the embryo's developmental stage, which is not identical to the embryo's chronological age. Our findings demonstrate that the period of 24 hr of in vitro development matches only a few hours of in vivo development.