Abstract
The histone deacetylase inhibitor (HDACi) has been investigated for treating cancers and many other diseases as well as enhancing the reprogramming efficiency in cloned embryos for decades. In the present study, we investigated the effects of two novel HDAC inhibitors, i.e., HDACi-14 and -79, at the concentrations of 0, 1, 2, or 4 μM on the development of embryos cloned by the oocyte bisection cloning technique (OBCT). Blastocyst rates for the reconstructed embryos reached 60% in the 2 μM HDACi-14-treated groups, which was higher (P < 0.05) compared to the untreated group (36.9%). Similarly, HDACi-79 treatment at 2 and 4 μM also conferred higher (P < 0.05) blastocyst rates than that of the untreated group (79.4, 74.2, and 50.0%, respectively). Both HDACi-14 and -79 treatments had no beneficial effect on total cell numbers and apoptotic indices of cloned embryos (P > 0.05). Histone acetylation profile by both HDACi-14 (2 μM) and -79 (2 μM) treatments demonstrated a drastic increase (P < 0.05) mainly in two-cell stage embryos when compared to the control group. After seeding on the feeder cells, the aggregated cloned blastocysts produced by the HDACi-79 treatment showed a significant increase of primary outgrowths compared to the control group (60.0% vs. 42.9%; P < 0.05). Finally, the cloned embryo-derived ES cell lines from aggregated cloned embryos produced from the HDACi-79-treated, HDACi-14-treated and control groups were established (5, 3, and 2 lines, respectively). In conclusion, the novel histone deacetylation inhibitors improve blastocyst formation and potentially increase the derivation efficiency of ES cell lines from the cloned porcine embryos produced in vitro. Depending on the purposes, some fine-tuning may be required to maximize its beneficial effects of these newly synthesized chemicals.
Highlights
Embryonic stem (ES) cells represent a population of pluripotent cell type capable of selfrenewal and differentiation into all somatic cell lineages
The cloned embryos were evaluated for their developmental competence after being respectively treated with each histone deacetylase inhibitor (HDACi), namely HDACi-14 or HDACi-79
When cloned embryos were treated with 2 μM HDACi-14, blastocyst rates were increased significantly (59.7%, P < 0.05; Table 2) compared to the Control (36.9%) and the 4-μM HDACi-14-treated groups (37.1%)
Summary
Embryonic stem (ES) cells represent a population of pluripotent cell type capable of selfrenewal and differentiation into all somatic cell lineages. ES or ES-like cells have been established from SCNT derived embryos in various species such as mice, rabbits, bovine, primates, and pigs [3,4,5,6,7,8,9]. The establishment of ES cell lines in domestic animals could potentially impact studies of mammalian embryogenesis and regenerative medicine. ES cells can be used to produce transgenic animals, such as pigs, serving as xenotransplantation donors. Pigs are a desirable species to create pluripotent cell lines because of their immunological, physiological and functional similarities to humans [10]. The morphology of porcine ES cell colony is similar to that of human ES cells which is more like flattened cell aggregates rather than dome-shaped colonies (as in murine species)
Published Version (Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have