ABSTRACT
BACKGROUND: In recent years the increasing use of nanoparticles has led researchers to study their effects on biological systems. The most important effects of nanoparticles on cells are their ability to induce or suppress production of reactive oxygen species (ROS). Changes in reactive oxygen species play an important role in various developmental processes, including proliferation and differentiation in several diseases such as Parkinson. The aim of this study was to investigate the effect of iron oxide nanoparticle with dimensions of less than 20 nanometers on the viability and neuronal differentiation of mouse embryonic stem cell (Royan B1). METHODS: To assess the effects of Fe2O3 nanoparticles on neuronal differentiation of Royan B1 cells, embryoid bodies were divided into eight groups receiving different amounts of nanoparticle (10, 20, 30 µg/mL) for 12 hours, retinoic acid (1 µM), and both. Differentiation was examined under phase contrast microscope and using immunocytochemistry. RESULTS: Data analysis showed that cell death was increased by a time and concentration manner and there was a direct relevance between iron oxide amount and H2O2 level in cells. Statistical analysis of embryoid bodies showed that neural differentiation of mouse embryonic stem cells in groups that received nanoparticles were significantly lower than other groups and their viability were considerably reduced. CONCLUSION: According to the findings of this study it can be concluded that iron oxide nanoparticles reduce retinoic acid-neuronal differentiation in mouse embryonic stem cells and it seems that the main mechanism involved in the reduction of viability and neural differentiation was enhanced levels of ROS within the cells.
Subject(s)
Antineoplastic Agents/administration & dosage , Embryonic Stem Cells/drug effects , Ferric Compounds/administration & dosage , Metal Nanoparticles/administration & dosage , Reactive Oxygen Species/metabolism , Tretinoin/administration & dosage , Animals , Cell Differentiation/drug effects , Dose-Response Relationship, Drug , Hydrogen Peroxide , Immunohistochemistry , Mice , Microscopy, Phase-Contrast , Neurons/drug effectsABSTRACT
There are growing numbers of recombinant proteins that have been expressed in milk. Thus one can consider the placement of any gene of interest under the control of the regulatory elements of a milk protein gene in a dairy farm animal. Among the transgene introducing techniques, only nuclear transfer (NT) allows 100 % efficiency and bypasses the mosaicism associated with counterpart techniques. In this study, in an attempt to produce a transgenic goat carrying the human coagulation factor IX (hFIX) transgene, goat fetal fibroblasts were electroporated with a linearized marker-free construct in which the transgene was juxtaposed to ß-casein promoter designed to secret the recombinant protein in goat milk. Two different lines of transfected cells were used as donors for NT to enucleated oocytes. Two transgenic goats were liveborn. DNA sequencing of the corresponding transgene locus confirmed authenticity of the cloning procedure and the complementary experiments on the whey demonstrated expression of human factor IX in the milk of transgenic goats. In conclusion, our study has provided the groundwork for a prosperous and promising approach for large-scale production and therapeutic application of hFIX expressed in transgenic goats.
