Volume 10, Nº 1 (2016)

A novel nonimmortalized fibroblast-like cell line SC6-MSC was derived from SC6 human embryonic stem cells (ESCs). Numerical and structural karyotypic analyses of these cells have revealed that it is hypodiploid: 45, X0. SC6-MSC average population doubling time was 26.0 ± 0.4 h at the 8th passage and 82.0 ± 9.2 h at the 18th passage. The growth curves showed active proliferation during passages 8–10 and consequent gradual decrease of the proliferative activity that stopped by the 20th passage. Flow cytometry analysis of surface markers has been carried to determine the line status. Revealed positive expression of CD44, CD73, CD90, CD105, and HLA-ABC and lack of CD34 and HLA-DR surface antigens are common for mesenchymal stem cells (MSCs). However, expression of CD90 and CD105 surface markers was significantly lower than for other MSC lines, including the line SC5-MSC derived from human ESC line SC5. Immunofluorescence analysis of surface markers and Oct-4 transcription factor, characteristic for human embryonic stem cells, showed the lack of Oct-4 expression and the presence of SSEA-4 and TRA-1-60 typical for a number of karyotypically normal MSC lines. Immunofluorescence assay showed the presence of the early differentiation markers of three germ layer derivates common for human ESCs. This shows that MSCs may be useful for reparation of tissue damage in corresponding microenvironments. It was revealed that SC6-MSC cells were able to differentiate into osteogenic and chondrogenic, but not adipogenic, directions. The results obtained indicate with high probability that disordered chromosomal and, accordingly, gene balance, in SC6-MSC line with karyotype 45, X0 result in decreased differential potential and expression of CD90 associated with the processes of cell differentiation and aging.

Human mesenchymal stem cells are a promising cell source for tissue engineering. During transplantation, they may be subjected to oxidative stress due to unfavorable cellular microenvironment characterized by an increased level of reactive oxygen species. Recently, we have demonstrated that oxidative stress response of human mesenchymal stem cells derived from endometrium (hMESCs) depends on the oxidizer concentration. The duration of cell treatment with an oxidizer also may play an important role. In this study, we investigated the dependence of the cell response on H₂O₂ treatment duration. The effects of high H₂O₂ doses on hMESCs and human lung embryonic fibroblasts were compared. In both cell types, H₂O₂ treatment for 60 min caused multiphase cell cycle arrest, with dose-dependent cell death occurring equally in all phases of the cell cycle. However, the cell death dynamics in hMESCs and fibroblasts were different. Interestingly, in both cell types, shortening of H₂O₂ treatment from 60 to 10 min induced growth retardation, G1-phase cell accumulation, and cell size increase. Collectively, these findings suggest that there is induction of premature senescence. Thus, shortening of oxidative stress induced in human endometrial stem cells and embryonic fibroblasts by high H₂O₂ doses enables one to modulate cellular response as both cell death and premature senescence.

The urokinase system, represented by a plasminogen activator of urokinase type (urokinase, uPA), urokinase receptor (uPAR), and inhibitors of plasminogen activator (PAI-1 and PAI-2), plays an important role in the regulation of vascular wall functioning. Urokinase signaling initiates proteolytic cascade and degradation of the extracellular matrix; and also activates intracellular signaling in vascular cells. This study is the first to reveal a urokinase-mediated fundamental mechanism that regulates the growth trajectory and branching morphogenesis of blood vessels. This mechanism may be of particular importance during vessel growth in early embryogenesis and in the adult during tissue regeneration.

The sex ratio in the first trimester of pregnancy shifts toward males due to increased elimination of female embryos. One reason for this phenomenon may be disruption of X chromosome inactivation. In this paper, we have analyzed the nature of the X chromosome inactivation in extraembryonic tissues of induced and spontaneous abortuses with 46,XX karyotype. Both equiprobable and asymmetric inactivation have been found in chorionic cytotrophoblast from spontaneous and induced abortuses. In the extraembryonic mesoderm of the control group of embryos, only equiprobable inactivation has been found, whereas this parameter was shifted in 15% of spontaneous abortions. The highest incidence of the selective inactivation of one of the parent homologues was found in the group with a lack of development of embryos and embryos from women with recurrent miscarriages. One of the reasons for the observed results can be compartmentalization of cells in the blastocyst leading to the nonrandom redistribution of cells and the predominance in the inner mass of cells with an active X chromosome with aberrations incompatible with normal embryonic development.

The polar growth of the pollen tube is a key stage in the life cycle of seed plants, which is critical for successful sexual reproduction. One of the most important components of this process is ion transport across the cell membrane coordinated in time and space. Different classes of signal molecules, including reactive oxygen species, as has been found recently, participate in regulation of ion transmembrane transport. In this study, based on the model system of subprotoplasts isolated from pollen tubes, we showed that hydrogen peroxide can regulate two targets located on the plasma membrane: nifedipine-sensitive Ca²⁺ channels and ion transport, both of which control the membrane potential. The interaction of hydrogen peroxide with these targets resulted in an increase in an intracellular Ca²⁺ concentration and hyperpolarization of the plasma membrane. Faster regeneration of the cell wall was a consequence of elevation of the Ca²⁺ intracellular concentration.

An optical tomography investigation of the nuclear cycle in large freshwater amoebae Amoeba proteus has been performed for the first time. Nuclei of cells from a synchronized culture were stained with DAPI and examined using a confocal laser scanning microscope. Detailed analysis of three-dimensional images of the intranuclear chromatin at different stages of the nuclear cycle has been performed. The materials obtained, in combination with the published data, allow for a completely new representation of the dynamics of the structural organization of the A. proteus nucleus during the cell cycle. Two-stage interphase and mitosis of a special type not matching any of the known types in the existing systems of classification of mitosis were found to occur in amoebae. Amplification of chromosomes and/or fragments thereof supposedly occurs during the cell cycle, which is consistent with the available data on nuclear DNA hyperreplication during the cell cycle of A. proteus. The number of chromosomes can vary at different stages of the cycle because of amplification, this being a putative reason for the discordant reports on the number of chromosomes in this species. The elimination of “excess” DNA mainly occurs during the transition from prophase to prometaphase. Finally, specific features of chromosome behavior during mitosis allow conclusion to be drawn that many, if not all, chromosomes are of a holocentric type.