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Vol 8, No 4 (2013)


Pamyati Georgiya Petrovicha Pinaeva

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Genes & Cells. 2013;8(4):6-7
pages 6-7 views

Exhaustion of endothelial reparation as factor of atherosclerosis progression

Khalimov Y.S., Babak A.V., Zatcepin V.V., Vetryakov O.V., Tcepkova G.A.


Atherosclerosis is a polyetiologic disease. Atherogenicdysli poproteinemiahas the greatest role in atherogenesis, but there are a lot of atherogenic factors independent of lipid profile. Contemporary scientific data underline the key importance of endothelial reparation in protection of endothelium from detrimental influences (smoking, dyslipoproteinemia, diabetes etc.).
Genes & Cells. 2013;8(4):29-32
pages 29-32 views

Methods of cells labeling for visualization in vivo

Solovieva A.O., Zubareva K.E., Poveschenko A.F., Nechaeva E.A., Konenkova V.I.


Today cell transplantation is promising treatment of various diseases. The therapeutic potential of cells depends on their migratory activity, the study of which is necessary for developing effective approaches of cell therapy. There are different approaches to investigate cells survival in vivo and spatial distribution in the organism after transplantation. This review summarizes the comparative characteristics of contrast agents such as fluorescent dyes, nanoparticles, radioisotope, reporter genes, and the features of their interaction with the cells. The main advantages and disadvantages of different methods of labeling cells have been analyzed.
Genes & Cells. 2013;8(4):33-38
pages 33-38 views

Phenotype and functional properties of adipose derived MMSC after interaction with cord blood mononuclear cells

Bobyleva P.I., Andrianova I.V., Maslova E.V., Andreeva E.R., Gogiya B.S., Buravkova L.B.


It is well known, that ex vivo expansion of hematopoietic stem cells is more effective when the culture conditions mimic their in vivo microenvironment. Multipotent mesenchymal stromal cells (MMSC) are widely used in such systems due to their ability to support hematopoiesis. In addition, the result of MMSC-hematopoietic cells' interaction can be affected by O 2 concentration, which is shown to modify the number of MMSC characteristics. The feedback effects of hematopoietic cells on MMSC during their interactions are less investigated. We studied the effects of cord blood mononuclear cells (MNC] and MMSC interaction on the later cells morphofunctional characteristics during 72 h coculture at 20% or 5% O 2. The dynamics of adhesion was estimated with CCD-camera equipped microscopy, the effects of the interaction with MNC on MMSC viability and CD54, CD44, CD90 expression were detected by flow cytometry. After interaction with MNC, MMSC displayed increased CD54 and in some cases - CD44 expression. The share of CD90+-MMSCs and their viability remained high during coculture with MNC at both O 2 levels. The adhesion of MNC was somewhat faster in 20% O 2. Thus, regardless of O 2 concentration coculture of allogenic MMSCs and MNC inc 2reased the expression of MMSC adhesion molecules involved in formation of specialized cell-to-cell contacts that provided effective adhesion of MNC. MMSC did not provoke allogenic MNC cytotoxic activity that is of particular importance in the case of potential therapeutic application of MMSC.
Genes & Cells. 2013;8(4):39-45
pages 39-45 views

Evaluation of apoptosis stages and posphatidylserine distribution in membrane of cord and peripheral blood nucleated cells at various cryopreservation protocols

Babijchuk L.A., Mykhailova O.O., Zubov P.M., Ryazantsev V.V.


Cryopreservation of cord blood nucleated cells for subsequent application in clinical practice is the only way of their long-term storage. The aim of this study was evaluation of the apoptosis stages and phosphatidylserine distribution in the nucleated cells membrane after cryopreservation by different methods. Nucleated cell fractions were frozen under the protection of the cryoprotectants with different mechanism of action. It was shown that nucleated cells isolation in polyglucin and subsequent freezing under 5% DMSO protection, as well as cells isolation by two-step centrifugation method and subsequent freezing under 10% of polyethyleneoxide (PEO) protection, allowed to keep intact most of the cord and peripheral blood cells. Cryopreservation of nucleated cells isolated using Ficoll, regardless of the used cryoprotectant, leads to significant disruption of the lipids asymmetric distribution in membrane and significantly reduces the number of living cells. It has been found that cord blood nucleated cells more resistant to damaging factors of cryopreservation than peripheral blood cells, that is shown in significant differences between the number of live cells in quite all cases before and after cryopreservation .
Genes & Cells. 2013;8(4):50-54
pages 50-54 views

Decellularization as a prevention of immune response activation to allogeneic pulmonary valves

Sergeevichev D.S., Sergeevicheva V.V., Subbotovskaya A.I., Vasilyeva M.B., Dokuchayeva A.A., Karaskov A.M., Kozlov V.A.


The top priority of clinical immunology is the study of alloreactivity mechanisms and development of methods for prevention of transplanted organs rejection. The discharge of vascular and valvular prostheses from a non-functional cellular debris is effective way to reduce their immunogenicity in the post-transplant period and to extend their functional usefulness. Obviously, removal of the maximum amount of HLA-determinants in cell membranes using decellularization should lead to reduction of complication after heart valves allogeneic transplantation. However, the key mechanisms of the immune response during acellular biomatrix implantation are insufficiently studied. In this study we investigated the influence of preimplantation treatment of human cardiac valves on the in vitro expression of important lymphocytes regulators CD28 and CD152 (CTLA-4). We found a significant rise expression of CD8+28+ from 37,9% to 44,5%, CD8+152+ from 0,3% to 22,6%, CD4+28+ from 96,4% to 98,4% and CD4+152+ from 0,04% to 43,8%, p<0,05 on lymphocytes after cultivation with native valves fragments and valves after cryopreservation, that indicated T-cells activation in the presence of cellular antigens. In contrast, no difference was found in the quantitative expression of CD28 and CD152 on T-lymphocytes after culturing with decellularized valves in comparison with the control, that indicating the absence of T-cell response to allogeneic acellular tissue matrix of the heart valves. Thus, from the point of applied immunology was shown that transplantation of decellularized allogeneic valve is preferable in comparison with native and cryopreserved grafts. Also identified candidate molecules of the transplantation immunity represent a possible therapeutic interest.
Genes & Cells. 2013;8(4):55-60
pages 55-60 views

Tissue-engineered construction made of adipose derived multipotent mesenchymal stromal cells, polylactide scaffolds and platelet gel

Buharova T.B., Volkov A.V., Antonov E.N., Vihrova E.B., Popova A.V., Popov V.K., Goldstein D.V.


We have shown the efficacy of tissue-engineered construction made of adipose derived multipotent mesenchymal stromal cells predifferentiated in the osteogenic direction, polylactide scaffolds fabricated by surface selective laser sintering, and platelet gel for bone tissue regeneration on the heterotopic osteogenesis model in rats.
Genes & Cells. 2013;8(4):61-68
pages 61-68 views

Sodium alginate bioresorbable membrane and octacalcium phosphate biomaterial for guided bone regeneration

Gunn A.N., Fedotov A.Y., Deev R.V., Komlev V.S.


To study the characteristics of a porous sodium alginate membrane and assess its role in guided bone regeneration of critical bone defects in rats' skulls containing ceramic octacalcium phosphate. The polymer membrane was made through lyophilization of 5 ml of 2% sodium alginate solution sealed with 10% solution of calcium/magnesium chloride (30/70). OCP granules were synthesized through processing calcium salts with orthophosphoric acid. The in vivo experiment involved 24 Vistar rats. Critical 9 mm diameter defects were made in their parietal bones. The animals were divided into four groups: 1st - control group, 2nd - the defect contained OCP granules and was covered with a skin flap, 3rd - OCP under an alginate membrane, 4th - membrane only without OCP. The animals were withdrawn from the experiment after one and three months. The membranes obtained had a porosity of up to 80% and a tensile strength of up to 2mPa. Ceramic OCP granules were porous, spherical, about 0.6 or 0.7 mm in size. XRF showed that the characteristic peak for OCP was at 4.8°0. During the in vivo experiment in the control group, the skin flap grew together with the dura mater and closed the defect. In the second group, OCP granules were allocated randomly in the soft tissues. In the third one, granules became organized within the bone defect and the alginate membrane was preventing the granules from migrating and the connective tissue from growing into the defect. Osteoid tissue was formed and then mineralized. In the fourth group, the alginate membrane gradually went disintegrating, with appearing solid parts resembling ossified fragments. No inflammation was observed. A study of the AM has shown that it matches all the requirements for barrier membranes, by being neutral to the adjacent tissues, not causing inflammation, being able to keep its structure for a long period, building solid, maybe mineralized fragments along with resorption areas, which is evidenced by the sealing lines, and not causing giant multinuclear cells to appear. OCP granules under the AM are gradually resorbed, building up osteoid tissue that then gets mineralized.
Genes & Cells. 2013;8(4):70-77
pages 70-77 views

Histogenesis of epithelial and stromal components at ductal adenocarcinoma of the pancreas

Setdikova G.R., Paklina O.V., Chekmareva I.A., Gordienko E.N.


The aim of our research is to define the morphological properties of anaplastic tumor site and surrounding stroma in ductal pancreatic cancer.The epithelial- stromal interactions in ductal pancreatic cancer cause its aggressive biological behavior, resistance to chemotherapy and survival of different depending on tumor differentiation. Heterogeneity of ductal carcinoma manifested in the appearance of anaplastic (sarcomatoid) component, which outlined the ability of epithelial tumor cells acquire the property of mesenchymal cells, which do not require the stroma and have aggressive malignant potential that influences the survival of patients .
Genes & Cells. 2013;8(4):78-81
pages 78-81 views

Evaluation of the quality and safety of cryopreserved human multipotent mesenchymal stromal cells derived from placenta for the clinical use

Astrelina T.A., Gomzyakov A.E., Kobzeva I.V., Karpova E.E., Kruglova A.Y., Scorobogatova E.V., Balashov D.N., Knyazev O.V., Yakovleva M.V.


The application of multipotent mesenchymal stromal cells (MMSC] in recent years is more widely evaluated in clinical trials. MMSC possess a number of unique features: the ability to support hematopoiesis, facilitating engraftment of HSC; immunomodulatory effects, including in regard to «graft versus host» reaction, and ability to differentiate into various cells types, that is defined their involvement in tissue and organ repair processes. The use of MMSC does not require antigenic compatibility control that leads to high potential accessibility of cell therapy. The most actively MMSC are evaluated in therapy of hematological, immunological, autoimmune, hereditary and other diseases. The aim of the study was to assess the quality and safety of cryopreserved placenta-derived MMSC prepared for clinical application. MMSC were obtained from 90 samples of placenta according to registered medical technology ФС №2010/374 от 13.10.2010. We evaluated the quality of placental MMSC by bacterial and viral control; determined the viability of cells with trypan blue and 7AAD; confirmed the immunophenotype specific for MMSC by flow cytometry. The biological safety of MMSC was analyzed by G-banding technique (15-20 metaphase chromosomes]. 72 samples of placenta-derived MMSC (193 doses] were prepared and used with HSC transplantation to 22 patients (42 MMSC doses] for optimization of HSC engraftment, prevention, control and treatment of «graft versus host» disease, liver regeneration. MMSC transfusion was performed at a dose of 2mln/kg. The results showed that allogeneic placenta-derived MMSC harvested in compliance with strict quality control were safe for clinical use and offered several significant advantages: no special invasive procedure for their tissue source explantation, high proliferative index on the 4th passage of cultivation, which allowed getting the required amount of cells for maximal therapeutic effect.
Genes & Cells. 2013;8(4):82-87
pages 82-87 views

Quantitative evaluation of the volume of the vascular bed in patients atherosclerotic lesions of lower extremities against therapeutic angiogenesis

Katelnitsky I.I., Katelnitsky I.I.


Results of treatment of 6 patients with obliterating atherosclerosis of the lower extremities by way of therapeutic angiogenesis - namely, the use of gennogi inducer of vascular growth of the drug «Neovaskulgen». The follow-up of more than three months. An original procedure scintigraphy by intra-arterial injection of Tc-phosphate complex pirfoteh quantify the capillary network of limbs prior to treatment and at 1 and 3 months after the introduction of the «Neovaskulgena». It is shown that the largest increase in the volume of the vascular bed made in the legs in place of the drug.
Genes & Cells. 2013;8(4):88-90
pages 88-90 views

Problems in the legal regulation of cord blood harvesting

Solonitsina L.A., Sazonov S.V., Leontiev S.L.


This work was aimed to analysis of the problems with medical activities organization related to the existing omissions in the Russian legislation regulating the use of cellular technologies. As a particular case, the cord and (or) placental blood harvesting was considered. In this article the current normative acts we analyzed in terms of regulation of cord/placental blood harvesting activity. The need to regulate this type of activity is caused by, on the one hand, present and growing demand in cord and (or) placental blood harvesting as the primary stage of its preparation for further storage of cellular material. On the other hand, it is necessary to provide the safety and quality of these medical services. Methods. We used juridical hermeneutics methods, the method of interpretation, and comparative jurisprudence. Results. The analysis of the current regulatory framework allows us to conclude that in the present cord and (or) placental blood harvesting has no proper legal regulation, which is a deterrent factor for cellular technologies development in Russia. Cord and (or) placental blood harvesting should be considered as a medical service and appropriate complex changes should be introduced in the normative acts to regulate and provide the accessibility and quality of medical service.
Genes & Cells. 2013;8(4):91-94
pages 91-94 views

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