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The article presents the present state of knowledge about obtaining in vitro embryos from farm animals. This biotechnique includes: in vitro maturation of oocytes, in vitro fertilizing matured oocytes and in vitro culture of embryos. The aim of in vitro production of embryos is to obtain more blastocysts and blastocysts of good quality which will determine the efficiency of embryo transfer and facilitate the production of a greater number of healthy offspring. Offspring were produced after transferring embryos produced in vitro in sheep, cattle, pigs, goats and horses. This biotechnique is used in farm animal breeding, biotechnology and basic research.
Surgically obtained early sheep embryos collected at morula, blastocyst, expanded blastocyst or hatching/ hatched blastocyst developmental stages were subjected to vitrification. The experiment was carried out by the traditional vitrification method, employing 0.25 ml insemination straws as carriers of embryos. The vitrification medium (VM) based on a modified phosphate-buffered solution (PB1) containing 3.0 g/L bovine serum albumin consisted of 1,2-propanediol, glycerol and sucrose (2.72 M, 1.36 M and 1.0 M - respectively). Embryos (each stage separately) were loaded into straws after 7-10 min of saturation in an equilibration solution (without sucrose). After 1 min period of saturation in VM embryos were plunged into liquid nitrogen. After warming, embryos were cultured in vitro and/or were surgically transferred to a surrogate mother for survival examination. Of 71 embryos tested, only 13 (18.3%) revealed development in vitro. The highest survival rate was observed in the most advanced group of embryos. Of 10 hatching/hatched embryos 6 (60%) survived (reexpanded). Three lambs were born after the transfer of 3 embryos from this group.
The aim of this review was to present the role of embryonic genome activation in zygote formation and in early embryonic development. Moreover, the authors emphasize the influence of selected factors on the quality and development of embryos in preimplantation stages. In recent years, reproductive biologists have focused on such processes as the regulation of oogenesis, folliculogenesis and morphogenesis. Thanks to the development of molecular biology and reproductive bio-techniques, it was possible to demonstrate the important role that activation of the embryonic genome plays in the above processes. Embryonic genome activation is a specific process whose origins dependent on the species of the mammal. In some species, activation of the embryonic genome begins in the 2-cell-stage embryo, while in others it begins during the 5th cell division. Molecular changes associated with embryonic genome activation play a crucial role in the morphological structure of the embryo. However, most of these morphological structure changes occur in the cell nucleus following the formation of nucleolus precursor bodies (NPB). It has also been suggested that epigenetic changes, such as the methylation and demethylation of embryonic DNA or the acetylation of histones, may play an important role in embryonic genome activation. There is little literature describing the influence of sperm RNA on basic semen parameters, the ability of a spermatozoon to fertilize an oocyte, or early mammalian embryonic development. This review discusses these parameters, as well as the role of micro-engineering and microfluidic research in the assessment of embryo quality.
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