Cultivation of mice embryos on various nutrient media up to the stage of two blastomers after the microinjection of a genetically-engineered construct

Abstract

The normal growth and differentiation of mammalian embryos in vitro during the pre-implantation period depends on the availability of appropriate metabolic substrates. Selection of optimal cultivation conditions when working with microinjected embryos is of significance, since any manipulations can reduce their viability. This is particularly important when obtaining transgenic animals. In order to determine the most effective culture medium, we studied the survival of mouse embryos after the microinjection of a genetically engineered construct into the pronuclei and further cultivation on the M16 and Onestep nutrient media. Compared to the M16 medium, an increase in the number of embryos that developed to the stage of two blastomeres on the Onestep medium was observed. Differences were also revealed in the two control groups, the embryos in which were not subjected to microinjection, but were cultured on the same media under the same conditions as the embryos of the experimental groups. We found that the Onestep medium, in which the percentage of survived embryos exceeded that in the M16 medium, is a more efficient medium for the cultivation of intact embryos.