No abstract

The advances in translational biomedical research, especially in genetic engineering, created new opportunities to trace the courses of human diseases and develop effective therapeutic methods. There remains, however, a growing demand for appropriate animal models for the precise evaluation of the efficacy and safety of new drugs or therapeutic concepts. Thus far, rodent models have been most widely used in translational research; however, since they do not perfectly reflect the human disease phenotype, transgenic pigs are increasingly being utilized as animal models. Thanks to the anatomical and physiological similarities between pigs and humans, swine are considered to be one of the most valuable animal models used in preclinical studies, including nutritional, metabolic and cardiovascular research. The resemblances involve the gastrointestinal, cardiovascular, urinary, respiratory, skeletal muscle and immune systems, as wells as body size, body composition and the omnivorous food choice. In addition, pigs are characterized by high fertility and fecundity, as well as the ease of use and low maintenance costs. Importantly, the existing efficient genetic engineering techniques enable relatively easy generation of tailored porcine models of human disease. One should be aware, however, of some physiological differences between humans and pigs to correctly interpret induced toxicological changes. The article provides an overview of current techniques for genetic modification of pigs, as well as the use of swine models in translational research exemplified by xenotransplantation, metabolic and coronary heart disease, and the gastrointestinal motility studies.

The aim of this study was to evaluate the effect of high hydrostatic pressure (HHP) on the in vitro developmental abilities of nano-transfected rabbit zygotes, their transfection efficiency, and the molecular quality of the blastocysts generated. This quality was assessed by estimating the quantitative profiles of Oct4, Casp7, and Bcl2 mRNA transcripts. The nano-transfection efficiencies of zygotes that had been pre-treated with either 20 MPa or 40 MPa of HHP (13.5% and 13.7%, respectively) were insignificantly lower than those found in zygotes not exposed to HHP prior to their nano-transfection (20.1%; P≥0.05). Moreover, applying HHP treatment with the parameters of 20 MPa and 40 MPa followed by the nano-transfection of zygotes brought about an insignificant decrease in the rates of embryos at the blastocyst stage (30.4% and 23.0%, respectively) as compared to the control group of nano-transfected zygotes (40.4%; P≥0.05). Furthermore, analyzing the transcriptional activity of Oct4, Bcl2, and Casp7 genes revealed that HHP enhances the relative abundance (RA) of all mRNA transcripts in blastocysts derived from non-transfected rabbit zygotes. In turn, the augmented RAs found in the pro-apoptotic Casp7 and anti-apoptotic Bcl-2 transcripts confirmed the onset and progression of programmed cell death in blastocysts developed from nano-transfected zygotes that had undergone HHP pre-treatment. The conceptualization based not only on a novel nano-transfection approach used to genetically modify in vivo-fertilized rabbit zygotes but also on their HHP pre-treatment is elaborated here for the first time, with an emphasis on further investigations aimed at producing transgenic rabbit and other mammalian species embryos by somatic cell cloning.