The activity of specific signaling and transcription factors determines the cell fate in normal development and in tumor transformation. The transcriptional profiles of gene-components of different branches of TGFbeta family signaling pathways were studied in experimental models of initial stages of three-dimensional in vitro differentiation of embryonic stem cells, embryonic germ cells and teratocarcinoma cells and in teratomas and teratocarcinomas developed after their transplantation into immunodeficient Nude mice. Gene profile analysis of studied cell systems have revealed that expression patterns of ActivinA, Nodal, Lefty1, Lefty2, TGF TGFbeta1, BMP4, and GDF were identical in pluripotent stem cells whereas the mRNAs of all examined genes with the exception of Inhibin betaA/ActivinA were detected in the teratocarcinoma cells. These results indicate that differential activity of signaling pathways of the TGFbeta family factors regulates pluripotent state maintenance and pluripotent stem cell differentiation into the progenitors of three germ layers and extraembryonic structures and that normal expression pattern of TGFbeta family factors is rearranged in embryonic teratocarcinoma cells during tumor growth in vitro and in vivo.

Ischemic stroke was modeled in white pedigreeless rats by the superficial blood vessel devascularization in the sensorimotor cortex. The preparations of neural progenitors--rat embryonic neural stem cells (rENSC) and human olfactory epithelium-derived neural stem cells (hOENSC) and differentiated fibroblasts ("cell control") were transplanted at the perimeter of the devascularized region. These cells marked with vital tracer stayed alive in the brain parenchyma for at least 16 days. The monitoring of contralateral forepaw motor deficit during 8 weeks demonstrated that only rats with rENSC transplantation had the stable and significant improvement of performance in cylinder test and swimming test (forepaw inhibition test) in comparison to "cell controls" and rats without cell transplantation. The maximal difference in the relative values (the efficacy) was 25% to the end of the experiment. There was no difference in the indicators of vibrissae-elicited forelimb placing test between experimental groups. The methodological approach used makes it possible to broaden the study of mechanisms of neural stem cells' therapeutic effect in stroke.

Aim of this study was elucidation of diagnostic value of qualitative and quantitative parameters of circulating endotheliocyte precursor cells (EPC), in the state of apoptosis (apoptotic endotheliocytes A AE), as well as effect of main risk factors on the studied parameters in patients with coronary atherosclerosis and hypercholesterolemia. We examined 24 patients with ischemic heart disease (14 men, 10 women, mean age 58.4 +/- 2.3 years) with stenoses >75% in coronary arteries according to angiography data and hypercholesterolemia (main group). Control group comprised 15 healthy volunteers. Circulating EPC in peripheral blood were evaluated by cytofluorimetry. Functional parameters of isolated EPC were assessed after cultivation in human fibronectin. Adhesive properties were studied in cell culture by immunehistochemical method after 7 days of incubation. Proliferation capacity was assessed by number of EPC in 1 mm3 of medium at day 7 of incubation. Determination and counting of AE was carried out using cytofluorimetry with immunostaining of cells by anexin B and CDA31 after preliminary treatment of endotheliocyte culture with H2O2 and comparing the results with control culture of endotheliocytes. Number of circulating EPC in patients with IHD was 58.% less than in healthy persons. Adhesive capacity of EPC in patients with IHD was lowered by 43%. Number of EPC surviving in cell culture was 80.9% less than in control group. At the same time number of circulating AE in patients with IHD was 2.5 times greater. Reduction of number of EDC, adhesive capacity, and rise of number of AE correlated significantly with number of involved coronary arteries, presence of diabetes, and smoking status. Presence of arterial hypertension and blood cholesterol level did not correlate with quantitative and qualitative parameters of EPC. Thus qualitative and qualitative parameters of circulating EPC and apoptotic cells can be considered as markers of dysfunction of endothelium and predictors of atherosclerotic vascular lesions in patients with hypercholesterolemia.

The aging process decreases the efficiency of the immune system is to maintain the constancy of the antigenic composition of the body. The process of recognition of own and foreign antigens is becoming less accurate, the intensity of immune responses is reduced. This leads to a characteristic of aging immunopathologic syndromes: an immune deficiency, autoimmunity, increased levels of circulating immune complexes, increased cases of benign monoclonal gammopathy. The weakening of the immune processes affected the clinical features of ulcerative colitis in the elderly. In old age, systemic manifestations of disease are observed in 2 times less, and local--in 2 times more likely than patients aged 20-40 years, with local complications such as profuse bleeding and a decrease in the lumen of the colon, there are 3-4 times more often. Thus, age is a factor that determines the characteristics of the course of ulcerative colitis, the severity of extra-systemic manifestations, prognosis and, consequently, response to therapy. Transplantation of mesenchymal stromal cells in elderly patients with ulcerative colitis is less effective than in patients with young and middle age.

To study the impact of modified nutritional support (NS) versus standard NS on therapy tolerability and posttransplantation in patients with oncohematological diseases.

The results of injection of the autologous stromal (mesenchymal) marrow-derived stem cells (ASMSC) transfected with green fluorescent protein (GFP) gene into the rat uterine scar were studied by methods of light microscopy. One week after ASMSC injection into the formed scar on the right side (2 months after the uterine horn ligation), large groups of blood vessels containing the formed elements of blood were found, while these were absent in the uterine scar on the opposite side. The study of the unstained sections in the reflected ultraviolet light demonstrated bright luminescence in the vessel endothelium and adventitia in the uterine horn scar only at the side of ASMSC injection. It is suggested that after the injection of ASMSC in the uterine horn scar, they form blood vessels as result of a differentiation into endotheliocytes and pericytes. The expression of GFP gene not only in vascularl endothelium, but also in their adventitia, indicated that the differentiation ASMSC is possible in both endotheliocytes and in pericytes.

The aim of the study was to determine the efficacy and safety of mesenchymal stromal cells (MSCs) of bone marrow in the treatment of patients with ulcerative colitis (UC).

Telomeres are the ends of the chromosomes and represent repeated DNA sequence and protein complex. Telomeres shorten with each cell division, which limits proliferative potential of cells. There is a great progress in clinical application of telomere length now. Additional characteristic of stem cell proliferation activity estimated by telomere length can be an informative indicator of transplant quality. In this work, we analyzed 14 cord blood samples by flow-FISH, ELISA and gematologycal analisator, and also the number of CD34 + CD45dim cells. Average telomere length of leukocytes fraction CB cells was 20.4-4.9 % respectively the control cells 1301 (T-cell lymphoblastic leukemia).

With the aim of formalization of gastric epithelium kinetics estimation the morphometric analysis of gastric mucosa bioptates was performed in 26 patients without endoscopic signs of stomach pathology. During step estimation of gastric pits at different depth authors analyzed changes of cells size, nucleocytoplasmic ratio, coefficient of nucleus elongation in immature cells of pit bottom (11 zone), differentiating cells in pit intermediate zone (L2), and on mucosa surface (L3 zone). Authors used claster and neuronet analysis (by Statistica and MedStat) to make the mathematical model which permit to determine the proportion of functional zones in normal pit (L--33 +/- 3%; L2--27 +/- 3%; L1--40 +/- 3% of gastric pit depth) and to develop the system of cell standards for each zone. These results can be used to diagnostic of gastric epithelium differentiation changes under pathology and to interpret the barrier properties of gastric epithelium.

A high-energy injury in subjects under the age of 50 as a rule leads to hip dislocation. This condition requires urgent orthopedic aid to avoid the risk of femur head osteonecrosis. Given a failure of closed reduction of dislocation (to be performed within 6 hours after the injury), immediate open reduction is indicated. The surgical approach depends on the dislocation type. The Kocher-Langenbeck approach is practiced to treat posterior dislocation and the Smith-Petersen or Watson-Jones approach in case of anterior dislocation. The most frequent and dangerous complications are femur head osteonecrosis (in 4-22% of the cases even after timely reduction) and post-traumatic coxarthrosis. Osteonecrosis is managed by a variety of surgical methods, viz. central decompresson, osteoplasty, corrective osteotomy, endoprosthetics. Its late stages, with sclerosis affecting over 30% of the head surface, are treated by resurfacing arthroplasty, hemiarthroplasty, total surface arthroplasty, uni- and bipolar hemiarthroplasty, total endoprosthetics, and administration of expanded stem cells into the subchondrial layer. The use of tissue-engineering technologies for the treatment of osteonecrosis is not confined to the treatment of its precollaptoid stages. They can be employed to promote osteointegration at the metal-bone interface and thereby to improve secondary stability or prolong the hardware lifetime.

This study demonstrated the possibility of in vivo activation of progenitor cells by hyaluronidase. Specifically, treatment with hyaluronidase increased the number of mesenchymal and bone marrow precursor cells, their proliferative activity, and differentiation. Also, it promoted stem cell mobilization into blood under effect of granulocyte colony-stimulating factor (G-CSF) and enhanced progenitor cell adhesive properties. Therapeutic efficiency of transplantation of mononuclear cells isolated from peripheral blood after administration of G-CSF improved under effect of hyaluronidase. Hyaluronidase immobilized by electron beam synthesis nanotechnology exhibited high specific activity with respect to stem cells via both enteral and parenteral routes.

The thanatogenetic mechanisms of stem cells (SC) of rapidly renewing system of intestinal epithelium still remain unclear. On the one hand, they are definitely involved into basic mechanisms of carcinogenesis in the gastrointestinal tract, because dysregulation of programs responsible for elimination of "unwanted" mutant cells (which are normally under immune and own intrinsic control) is one of the reasons of neoplastic expansion. On the other hand, elucidation and characterization of the regulatory machinery controlling SC survival are interrelated with problems of clinical medicine, including the increase of therapeutic efficiency of treatment of inflammatory and ulcer lesions of the gut, traumatic and surgical wounds, as well as restriction of side effects in normal tissues induced by application of intensive methods chemo- and radiotherapy of cancer. The latter is especially important for treatment of blood diseases and tumors of peritoneal cavity organs mainly due to bone marrow and intestinal epithelium damage. (These tissues are the most sensitive to these treatments.) The review considers data on exogenous and genetic modifiers of SC survival, and also the basic principles of mechanisms involved into renewal and regeneration of SC and the effects of microbiota on these processes.

Human embrional stem cells (hESC) are able to maintain pluripotency in culture, to proliferate indefinitely and to differentiate into any somatic cell type. Due to these unique properties, hESC may become an exceptional source of tissues for transplantation and have great potential for the therapy of incurable diseases. Here, we review new developments in the area of embrional stem cells and discuss major challenges--standartization of protocols for cell derivation and cultivation, identification of specific molecular markers, development of new aprouches for directed differentiation etc.--which remain to be settled, prior to safe and successful clinical application of stem cells. We appraise several potential approaches of hESC therapy including derivation of autologous cells via therapeutic cloning (1), generation of immune tolerance to allogenic donor cells via hematopoetic chimerism (2), and development of the banks of hESC lines (3). In addition, we discuss brifly induced pluripotent cells, which are derived via genetic modification of autologous somatic cells and are analogous to ESC. Our analysis demonstrates that uncontrollable differentiation in vivo and teratogenic potential of hESC are critical limitations of their application in clinic. Therefore, the major direction of hESC use is derivation of a specific differentiated progeny, which has lower proliferative potential and immune privilege, yet poses fewer risks. Finally, cell therapy is far more complex and resource-consuming process as compared to drug-based medicine; pluripotent stem cell biology and technology is in need of further investigation and development before these cells can be used in clinics.

Professor Yu.M. Lopukhin has made the most important contribution to the development of bioethics in this country based on his extensive experience in research, education and organization pertaining to different biomedical disciplines and high-tech medicine. The main works of Yu.M. Lopukhin on major bioethical problems of special importance for present-day Russia are analysed including ethical expertise of biomedical research and their legal aspects including tissue and organ transplantation, the use of embrionic stem cells, etc. The importance of international collaboration in the field of bioethics is emphasized.

The article describes successful use of allogeneic mesenchymal stem cells in complex treatment of patients with enzyme depended ulcerative colitis.

Recently we found that transposition of homologous chromosomes 1q12 loci towards the nuclear centre and activation of the chromosomal nucleolus-forming regions (NFR) are observed in human lymphocytes after exposure to low doses of X-radiation (10 cGy). These cell reactions were studied for human breast cancer stem cell cultures. There are two cell types in cell culture from single donor: with two (type 1) and three (type 2) loci of 1q12. It was shown that an adaptive response induced by X-ray irradiation is developed only in cells of the type 1 but not in type 2 ones after 3 and 10 cGy doses. We observed a considerable death of cell type 2 after low-dose exposure. Activation of the NFR in breast cancer stem cells after irradiation was not found. In this paper we discuss features of studied cancer stem cells lines and their responses to low doses of ionizing radiation.

Motor dysfunctions in Parkinson's disease are believed to be primarily due to the degeneration of dopaminergic neurons located in the substantia nigra pars compacta. Numerous cell replacement therapy approaches have been developed and tested, including these based on donor cell transplantation (embryonic and adult tissue-derived), adult mesenchymal stem cells (hMSCs)-, neural stem cells (hNSCs)- and finally human embryonic stem cells (hESCs)-based. Despite the progress achieved, numerous difficulties prevent wider practical application of stem cell-based therapy approaches for the treatment of Parkinson's disease. Among the latter, ethical, safety and technical issues stand out. Current series of reviews (Cell therapy for Parkinson's disease: I. Embryonic and adult donor tissue-based applications; II. Adult stem cell-based applications; III. Neonatal, fetal and embryonic stem cell-based applications; IV. Risks and future trends) aims providing a balanced and updated view on various issues associated with cell types (including stem cells) in regards to their potential in the treatment of Parkinson's disease. Essential features of the individual cell subtypes, principles of available cell handling protocols, transplantation, and safety issues are discussed extensively.

We suggested that satellite proliferation with subsequent incorporation of their nuclei in the fibres was essential for enhancement of the protein synthesis in the rat relieved m. soleus. In passive stretch of the muscle, the number of labelled cells exceeded 2.5-fold their number in hung out animals and 1.7-fold in the control animals. The expression level of the insulin-like growth factor I (IF-1) was determined in the m. soleus after hanging out with stretching, and no changes were revealed as compared with hanging out without stretching in the control group. We believe that the muscle IF-1 and satellite cell incorporation are not essential for preventing atrophy of relieved soleus muscle in stretching.

The changes in the state of long-term culture of blastocysts derived from female C57BL/6 mice after crossing with C57BL/6-Tg(ACTB-EGFP)1Osb/J males with a green fluorescent protein transgene EGFP on chromosome 15 were studied. Possible causes of different culture results were analyzed: the preservation of undifferentiated cells as dense clusters in the inner cell mass or their differentiation into extraembryonic endoderm. Comparison of the events going in blastocysts with the -/- or -/EGFP genotypes demonstrated that the GFP presence has no effect on cell processes. This allows us to use embryonic material from this mouse line in experiments that require long-term vital observation of embryonic cells.