The current research was conducted to explore thein vitrodevelopmental outcome and cytological/molecular quality of porcine nuclear-transferred (NT) embryos reconstituted with adult bone marrow-derived mesenchymal stem cells (ABM-MSCs) that were epigenetically transformed by treatment with nonspecific inhibitor of histone deacetylases, known as trichostatin A (TSA). The cytological quality of cloned blastocysts was assessed by estimation of the total cells number (TCN) and apoptotic index. Their molecular quality was evaluated by real-time PCR-mediated quantification of gene transcripts for pluripotency- and multipotent stemness-related markers (Oct4, Nanog, andNestin). The morula and blastocyst formation rates of NT embryos derived from ABM-MSCs undergoing TSA treatment were significantly higher than in the TSA-unexposed group. Moreover, the NT blastocysts generated using TSA-treated ABM-MSCs exhibited significantly higher TCN and increased pluripotency extent measured with relative abundance ofOct4andNanogmRNAs as compared to the TSA-untreated group. Altogether, the improvements in morula/blastocyst yields and quality of cloned pig embryos seem to arise from enhanced abilities for promotion of correct epigenetic reprogramming of TSA-exposed ABM-MSC nuclei in a cytoplasm of reconstructed oocytes. To our knowledge, we are the first to report the successful production of mammalian high-quality NT blastocysts using TSA-dependent epigenomic modulation of ABM-MSCs.

A broad spectrum of cytologically specific immunophenotypic, biochemical, physiological and molecular attributes of neonatal and adult mesenchymal stem cells (MSCs) exists. Taking into account the biological traits of these types of multipotent stem cells, they play a pivotal role in the wide utilization of MSC-mediated engineering and MSC-based assisted reproductive technologies (ARTs) for animal production, veterinary research, regenerative and reconstructive medicine, and cell/tissue transplantology. Consequently, this paper sought to comprehensively provide research highlights, and insightful interpretation of current goals and future targets related to the agricultural, embryological and biomedical application of MSC-assisted biotechnological strategies and MSC-dependent interdisciplinary research techniques.

Regulation of gene expression is a complex process. Epigenetics is the study of heritable changes in gene expression independently of DNA sequence. Epigenetic control of gene transcription is based on two main processes. The first is reversible DNA methylation, primarily of cytosine at position C5, rarely in position N3, or of adenine at position C6 (Xu et al., 2010). The second process is the change in chromatin structure and function by chemical modification of histones, including mainly methylation, acetylation, and phosphorylation of histone amino acids (Zamudio et al., 2008). During development and differentiation of cells, changes occur in DNA methylation of genes. After fertilization there are dynamic histone modifications and changes in DNA methylation in zygotes. Use of methylation sensitive restriction enzymes causes a global demethylation in the early embryonic stage (Sulewska et al., 2007 b). De novo methylation of CpG sites is followed by embryo implantation. Next, during gastrulation most genes are methylated except the tissue-specific genes. The last wave of de novo methylation takes place during the gametogenesis and is dependent on sex (Sulewska et al., 2007 b). The aim of this work is to review the current knowledge about epigenetic mechanism of molecular changes in animal cells with particular regard to embryonic development.

The aim of this study was to investigate the impact of the source of mesenchymal stem cells (MSCs) and their adipogenic derivatives on the relative abundances (RAs) noticed for mRNA transcripts of the selected adipokines (adiponectin and leptin) and their receptors. MSCs were isolated from bone marrow (BM) and subcutaneous adipose tissue (AT ) samples collected post mortem from a total of four gilts (each at the age of 6 months). The stemness of the MSCs was proven via recognizing their abilities to differentiate into adipocytes, osteoblasts, and chondrocytes. By using real-time PCR (RT -qPCR), the quantitative levels of transcriptional activity pinpointed for the adiponectin ( ADIPOQ ), adiponectin receptor 1 ( ADIPOR1) , adiponectin receptor 2 ( ADIPOR2) , leptin ( LEP ), and leptin receptor ( LEPR ) genes were comparatively analyzed between non-differentiated AT -MSCs and BM-MSCs and their cell counterparts undergoing differentiation into adipocytes. A significantly higher RA for ADIPOR1 transcripts was identified in the AT-MSCs compared to the BM-MSCs and their adipogenic derivatives (P≤0.05). Moreover, the quantitative levels of LEPR transcripts were shown to increase significantly among adipocytes originating from differentiated BM-MSCs compared to undifferentiated BM-MSCs, AT -MSCs, and AT -MSC-derived adipocytes (P≤0.05). Nonetheless, there was no significant inter-group variability in the RAs of the ADIPOQ , ADIPOR2 , or LEP mRNA transcripts (P>0.05). Cumulatively, the in vitro models focused on the identification and detailed exploration of transcriptomic signatures of undifferentiated pig BM- and AT -MSCs, and the molecular mechanisms underlying the adipogenic differentiation pathways of the above-indicated two distinct sources of stem cells were developed and optimized in the current investigation for the very first time. These ex vivo porcine models might confirm the expedited functional mobilization of ex situ -protected MSCs and their enhanced capacity to be transcriptionally reprogrammed into adipocytes due to physiopathological alterations in the expression profiles of adipokines and their receptors, which are prompted and progressed in obese and superobese women patients at the peri-pubertal period of ontogenesis. Finally, the models providing comprehensive molecular insights into ex situ -protected porcine BM- and AT -derived MSCs and their differentiated derivatives can be largely suitable for the biotechnologically assisted rescue and restitution of endangered representatives of rare native breeds of domestic pig.