Endoglin (CD105) is the marker of endothelial and mesenchymal stem cells and the component of TGF-β, BMP-9 and BMP-10-binding receptor complexes. Its expression is significantly increased on blood vessels endothelium of ischemic tissues and growing tumors. Measurement of concentration of the soluble endoglin in the serum or urine is used as a method for diagnosing cancer and pregnancy disorders. The aim of this work was to create a novel family of monoclonal antibodies recognizing endoglin on the cell surface and in biological fluids. Murine myeloma cells' derived recombinant protein representing the whole extracellular part of endoglin was used as an antigen. F1(SJL/JxBALB/c) mice were the donors of immune splenocytes. Hybridoma screening procedures were performed using E. coli-produced copies of the antigen, endoglin-expressing immortalized human cell lines, and primary cultures of human mesenchymal stromal cells. Ten novel monoclonal antibodies recognizing at least eight distinct epitopes were produced. Eight antibodies bind membrane form of endoglin on the surface of normal and transformed human cells derived from different tissue sources. Two antibodies recognize linear antigenic determinants of the molecule and can be used to detect endoglin by western blot. Sandwich ELISA system was designed in order to measure soluble endoglin in cell culture medium.

Cellular spheroids were derived from mesenchymal stem cell lines derived from 5-6-weeks embryo from different tissues of 5-6-week human embryo: bone marrow (FetMSC) and muscle of limb (M-FetMSC). Comparative analysis of the characteristics of these lines has been performed with 2D culturing in monolayer and 3D culturing in spheroids. The characteristics of cellular spheroids were obtained after 48 h after their formation from monolayer cultures on the 6th passage after decryopreservation. Spheroids in contrast to monolayer cultures are heterogeneous cell populations composed of fibroblast-like and epithelioid cells. Two-day spheroids are actively proliferating structure. Cell surface markers were analyzed using flow cytometry. Both in the monolayer cultures and cellular spheroids, this analysis has revealed the presence of expression of surface antigens CDD44, CD73, CD9O, CD105, HLA-ABC that are characteristic of human MSC, and the absence of expression if CD34 and HLA-DR. Nevertheless, the level of expression of CD90 and CD105 antigens was significantly lower in the spheroids as compared with corresponding monolayer cultures. Immunofluorescence and flow cytometry analysis of the expression of transcriptions factors and surface antigens characteristic of human embryonic stem cells showed the presence of expression of Sox-2 and SSEA-4 in 2D and 3D cultures. Lack of expression of Oct-4 in 2D cultures and its significant increase in 3D cultures has been found. Immunofluorescence analysis showed the presence of the markers of early differentiation in the derivates of three germ layers characteristic of human embryonic stem cells in the cellular spheroids of both lines, which coincides with 2D cultures of these lines. The directed osteogenic, chondrogenic and adipogenic differentiation of these lines has been shown. However, a number of differences has been found between monolayer cultures and spheroids. Adipogenic differentiation was more active in the cellular spheroids from cell line M-FetMSC a compared with corresponding monolayer cultures. Differences between the 2D and 3D cultures of both lines have been shown by the character of chondrogenic differentiation. The results obtained confirm the status of MSC for the cellular spheroids derived from monolayer cultured of cell lines FetMSC and M-FetMSC and apparently indicate a partial extension of their differentiation capacity as compared to monolayer cultured.

Musculoskeletal functions disorders may develop as a consequence of injuries and various types of congenital / acquired diseases, among which a special place belongs to muscular dystrophy. The technology with use of cells possessing myogenic potential is considered as one of the most promising approaches to solve the problem of effective restoration of skeletal muscles structure and function. In part I of the article the characteristic features, functions and phenotypic characteristics of satellite cells (SC) are reviewed as key factors of skeletal muscle tissue regeneration. Presented analysis of research results (preclinical and clinical) concerning therapeutic possibilities of technology using SC. In the second part of review will be presented data of the therapeutic use of stem cells of muscle and non-muscle origin for the treatment of skeletal muscles diseases.

Experiment was evaluated on 40 male Wistar rats. On the experimental model of mandible injury, bone autologous graft from tibia was placed on the surface of mandible (host bone). In the main experimental group, consisting of 20 animals, autologous rat adipose-derived stromal cells (ADSCs) were inoculated in space between autograph and host bones. ADSCs were not inoculated in the group of comparison. In experimental group with inoculated cells, the formation of a new fibroreticular bone structures in space between autograph and host bone was observed. These structures further underwent secondary reorganization and differentiation during the process of remodeling. As a result of the conducted study it was shown that in the experimental group by the day 180, statistically significant reduction of the area occupied by an immature fibroreticular bone took place. The reported phenomenon could be explained as a result of decline of the number of active cells in the population of inoculated ADSC, which is in consent with theory of limited cell division number due to telomeres shortening, described by Hayflick L. and Moorhead P.S. (1961).

The paper analyses the morphological and functional features of cord blood cells (CBCs) in the co-culture with multipotent mesenchymal stromal cells (MMSCs) from human adipose tissue under tissue-related oxygen. We have established that MMSCs effectively maintained viability of CBCs at different oxygen concentrations (1, 5 and 20%). According to the data obtained, the oxygen concentration affected the number of colony-forming units (CFUs) formed by CBCs in selective medium. In particular, not co-cultured CBCs the 1 and 5% O2 than at 20% O2. After co-culture with MMSCs, the CFUs numbers were similar at 1 and 20% O2 and increased by 30 at 5% O2. Tissue related O2 concentrations had an impact on the proportion of lineage-resctricted CFCs among CBCs: the number of more committed progenitors--CFU-G and CFU-M, increased, and multi and bipotent--CFU-GEMM and CFU-GM, decreased at low oxygen concentrations. This effect was more pronounced after co-culture compared to that of initial CBCs. Thus, the presence of stromal cells and tissue-related oxygen jointly and severally influenced CBCs in vitro.

Evaluate the fraction of various TLR1-TLR6 haplotypes in populations of Russians Bashkir and Nagaybak of Chelyabinsk Region.

This rare type of acute leukemia, blast cells of which express myeloid and/or lymphoid markers, is mainly diagnosed using flow cytometric findings. The paper describes a clinical case of mixed-phenotype acute leukemia, in which B-cell lymphoid antigen expressions were revealed by a flow cytometric technique, while bone marrow morphological specimens showed the signs of myeloid differentiation specific to blast cells. It is concluded that there is a need for a comprehensive examination of patients with new-onset acute leukemia and for an aggregate analysis of flow cytometric results with morphological and cytochemical findings.

Using a TV device to study microcirculation in brain we found that intracerebral transplantation of mesenchymal stem cells to 12-months old rats led to a significant increase (circa 1,5-fold times) of microvascular density in pia tissue and to increased constriction reactions of pia arterioles in response to noradrenalin application on a brain surface. Both microvascular density and pia arterioles reactivity was completely preserved in aging until 22-24 months.

Previously we've described the obtainment of a subpopulation of cancer stem cells from a human colorec- tal carcinoma cell line MIP101. These cells possess elevated clonogenic and tumorigenic capacities. According to our data, depletion of stem compartment in a cancer cell population blocks its tumorigenicity. The current work is dedicated to the comparison of tumorigenic potential between cell populations with enriched or depleted stem compartment. We show that tumor growth following xenografting of enriched stem cell population can be suppressed by intramuscular injections of ganciclovir. Thus, we report a method to obtain a cell population with high Oct4 promoter expression within the MIP101 colorectal carcinoma cell line and to eliminate these cells from the population in vitro as well as in vivo.

It is shown that the effect of 5-fluorouracil (5-FU) on mesenchymal stromal cells (MSCs) of the bone marrow and fetal liver of the rat increases the population of cells with a reduced proliferative potential. It is shown that, in the course of passaging, the 5-FU-resistant MSCs of the bone marrow and fetal liver lose their ability to differentiate in the osteogenic direction more and less rapidly, respectively, than the 5-FU-sensitive MSCs.

Brain stem cells represent an extremely intriguing phenomenon. The aim of our review is to present an integrity vision of their role in the brain of mammals and humans, and their clinical perspectives. Over last two decades, investigations of biology of the neural stem cells produced significant changes in general knowledge about the processes of development and functioning of the brain. Researches on the cellular and molecular mechanisms of NSC differentiation and behavior led to new understanding of their involvement in learning and memory. In the regenerative medicine, original therapeutic approaches to neurodegenerative brain diseases have been elaborated due to fundamental achievements in this field. They are based on specific regenerative potential of neural stem cells and progenitor cells, which possess the ability to replace dead cells and express crucially significant biologically active factors that are missing in the pathological brain. For the needs of cell substitution therapy in the neural diseases, adequate methods of maintaining stem cells in culture and their differentiation into different types of neurons and glial cells, have been developed currently. The success of modern cellular technologies has significantly expanded the range of cells used for cell therapy. The near future may bring new perspective and distinct progress in brain cell therapy due to optimizing the cells types most promising for medical needs.

In this review, we systematized the data that characterize phenotypic properties and functional features of T- and B-lymphocytes of immune memory. We examin the organization of T-cells of immune memory in a population, and their selective distribution in the organism according to their effector potential.

Experiment was conducted on 40 rats of Wister line. On the artificially reproduced experimental model autogenic mesenchimal stem cells (MSC) of adipose tissue were inoculated in space between bone autograph of tibia and mandible. MSC wasn't inoculated in the comparison group. Formation of a new bone substance in space between an autograph and mandible bone was observed. It was clear that after 120 days (180 days), there was a statistically significant decline of the area occupied by an immature fibroreticular bone. Described phenomenon, presumably, could be explained as a result of decline of the number of active cells in the population of inoculated MSC according to phenomenon of limited number divisions of cells on telomeres, described by Hayflick L. and Moorhead P.S.

The study of the immune system of mice of CBA (n=7) and C57BL/6 (n=7) strains was performed during a 12-hour photoperiod (12:12) and round-the-clock illumination (RI) in December and March. The numbers of CD3+, CD4+8-, CD4-8-, CD4+8+, CD19+, CD3+hi lymphocyte subpopulations in the thymus and spleen, as well as the proliferative activity of thymocytes and splenocytes were measured by flow cytometry. It was found that the most pronounced. interstrain differences in the cellular composition of the thymus were observed in March, while those in the cellular composition of the spleen were detected in December. Overall, in C57BL/6 mice as compared with CBA mice, the number and proliferative activity of the cells of lymphoid organs was increased. In December, the influence of RI on the cellular composition of the thymus was more pronounced in C57BL/6 mice, while the effect on cellular composition of the spleen was greater in CBA nice. In March, after exposure to RI, a number of CD3+, CD19+ and CD4+8-splenocytes was increased in CBA mice, while it was decreased in C57BL/6 mice. In C57BL/6 mice, after exposure to RI, the proliferative activity of thymocytes decreased in winter and increased in spring . Interstrain and seasonal differences in the cellular composition of lymphoid organs in CBA and C57BL/6 mice, with a photoperiod of 12:12 and RI should be taken into account in experimental work with aninials of different genotypes in different seasons of the year.

The review summarizes current knowledge on the Hedgehog signaling pathway, its role in normal embryogenesis and/or initiation and progression of neuro-oncological diseases, especially of high-grade gliomas, the most malignant neuroepithelial tumors. The main proteins forming the Hedgehog signaling pathway include Shh, PTCH1, SMO, HHIP, SUFU and GLI1 isoforms. Effects of other signaling pathways on the family of transcription factors GLI and other proteins are described. The review summarizes modern data about the impact of the Hedgehog signaling pathway on proliferation, migration activity and invasiveness, and also on tumor neoangiogenesis and tumor cell chemoresistance. The role of the Hedgehog signaling pathway in origin of cancer stem cells and epithelial-mesenchymal transition is also analyzed. Some prospects for new anticancer drugs acting on components of the Hedgehog signaling pathway inhibitors are demonstrated.

Cancer-testis antigens are expressed in the spermatogenic and cancer cells as well as in human and mouse pluripotent stem cells. However, the role of cancer-testis antigens of Mage families in the regulation of cellular processes in embryonic cells is largely unknown. In the present study comparative quantitative analysis of the gene expression of Magea and Mageb families was performed in mouse embryonic somatic cells (mouse embryonic fibroblasts, MEFs), long-term cultured in vitro and exposed to factors that inhibit and stimulate proliferation. The analysis revealed low expression of cancer-testis antigens of Mage families and showed that a lower proliferative activity of MEF at late passages was accompanied by slight up-regulation of the Magea gene expression and down-regutation of Mageb gene expression. However, modulation of the activity of MEK/ERK-signaling pathway and DNA demethylation by 5-azacytidine had no significant effects on the expression of Magea and Mageb genes in M EFs. The most essential changes in the expression levels of Mageb and Magea genes were found only when MEFs were exposed to mitomycin C. In all experimental variants, predominantly cytoplasmic localization of Mage antigens was found in MEFs at the phase of DNA synthesis, as well as at other cell cycle phases. Presumably, in actively proliferating mouse embryonic somatic cells the antigens of Magea and Mageb families can act as co-activators in the regulation of cell proliferation and other cellular functions.

This review is dedicated to comparative analysis of the early stages of thymus ontogeny in fish, amphibians, and mammals. Morphological and molecular-genetic aspects of the formation of thymic stroma, colonization of this organ with T-cell progenitors, and interaction of different cell populations in the course of organogenesis are considered. Particular attention is given to the hematopoietic role of the thymus during embryogenesis and new data on the origin of T-cell progenitors. The hypothesis about the possible presence in the organ of a self-sustaining population of stem cells, formed regardless of fetal hematopoiesis areas, is discussed.

Possibility of using stem cells and developing tissue engineering for regeneration of jaws bone defects, bones vault of the skull, dental and periodontal tissue are presented in the article.

Gene function disclosure and the development of modern technologies of genetic manipulations offered the possibility of genetic reprogramming application to alter cell specialization. With the involvement of a gene set that encodes the transcription factors responsible for the pluripotent state, any cell of an adult body could be reprogrammed into the embryonal.state and pluripotency could be induced in this cell. Such reprogrammed cells were called induced pluripotent stem cells (iPSCs), and they are capable of again passing through all developmental stages. This provides new possibilities for studies of the basic mechanisms of developmental biology, the formation of specific cell types, and the whole body. In culture, iPSCs could be maintained permanently in a nontransformed state and permit genetic manipulations while maintaining their pluripotent properties. Such a unique combination of their properties makes them an attractive tool for studies of various pathologies and for the delineation of treatment approaches. This review discusses the basic and applied aspects of iPSCs biology.

Short (25-35 nucleotides) regulatory piPHK, along with RNA-binding proteins of the Piwi family, constitute an evolutionarily conserved system that functions mainly in eukaryotic gonads. The system can be regarded as a variant of the mechanism of RNA interference, which is based on the recognition of target RNA as a result of complementary interactions with piRNA. The variants of this regulatory system function in the germline cells, including stem cells and somatic cells of the niche, ensuring maintenance of the germline stem cells and their differentiation. One of the most important functions (but not the only one) of this system is the repression of transposons, which guarantees genome stability in germline cells. This review focuses on the works of the authors of the review in the context of outstanding international achievements in a rapidly evolving re- search area, the biology of piRNA and the function of the Piwi protein.