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Vol 7, No 1 (2012)

Articles

OT REDAKTsII

OT REDAKTsII -.
Genes & Cells. 2012;7(1):6-22
pages 6-22 views

Mesenchymalstem cells from various human tissues: biological properties, assessmentof quality and safetyfor clinical use

Shachpazyan N.R., Astrelina T.A., Yakovleva M.V.

Abstract

Developing direction cellular medicine is the use of the unique properties of progenitor cells with high biological activity and potential for differentiation. Multipotent mesenchymal stromal cells (MMSC) are pluripotent stem cells, has several important properties for clinical use. The use of MMSC cultured «ex vivo» opens the question about the quality and safety culture for clinical use. The purpose of this review was to develop a program of cultivation and study of important properties of samples of human MMSC for clinical application. The review shows the characteristic phases of assessing the quality and safety of MMSC, including the cultivation of cells «ex vivo», immunological assessment, growth, immunomodulatory, and regenerative properties of the progenitor, an assessment of genetic and microbiological safety. Analyzed the assessment the quality and safety of MMSC «in vitro» tests. It is shown that the severity of the properties of each samples different and depends on the source and culture conditions MMSC.
Genes & Cells. 2012;7(1):23-33
pages 23-33 views

Heart valve tissue engineering: decellularization of allo- and xenografts

Kurapeev D.I., Lavreshin A.V., Anisimov S.V.

Abstract

Implantation of artifitial heart valve prostheses is assotiated with several serious complications. Therefore, there is a need in new approaches of preparing heart valve prostheses. One of such approaches is heart valve tissue engineering. Tissue engineered heart valves are to be biocompatible, durable, long-life, no necessity in anti-trombotic therapy, and they have potential to grow and regenerate with host. The most developed method of heart valve tissue engineering is decellularization, i.e. preparing an extracellular matrix, which has potential in recellularize with stem host-cells and then implant it. This review focuses on methods of decellularization of heart valves and their ability to change structural and biomechanical properties of allo- and xenografts.
Genes & Cells. 2012;7(1):34-39
pages 34-39 views

In vitro expansion and lineage commitment of the human umbilical cord blood myeloid progenitors

Petyovka N.V., Goncharova N.V., Seviaryn I.M., Kosmacheva S.M., Potapnev M.P.

Abstract

In vitro proliferation and myeloid differentiation of human umbilical cord blood (UCB) CD34+ cells with/without bone marrow multipotential mesenchymal stromal cells (BMMMSCs) feeding layer in serum-free medium supplemented with IL-3, IL-6, SCF and Flt3/l were investigated. Mononuclear cells (MCs) were isolated from the healthy donor UCB specimens (n = 4), CD34-enriched MCs were obtained by positive immunomagnetic selection (CD34+-rich). Both cell fractions (MCs and CD34+-rich) were expanded for two weeks in vitro. Cells were seeded to the new vessels when necessary. The percentage of CD34/CD45-positive cells was accounted by flow cytometry, subpopulations of myeloid progenitors were assessed in colony-forming tests. Upon two weeks of culture the number of CD34+ cells in MC fraction augmented by 70±29 and 980±414-fold with and without BM-MMSC feeding layer respectively and was not significantly differed from that in CD34+-rich fraction. Progenitor cell subpopulations were not altered during the first week of culture but granulocytopoiesis was superior over erythropoiesis during the second week. CD34 and CD45 expression patterns revealed two hematopoietic cell populations, one of them differentiating on the first week of expansion, and the other - on the second one. BM-MMSC feeder promoted significantly higher (p < 0,05) expansion rates of CD34+ cells and myeloid progenitors in comparison with non-feeding culture. BM-MMSC feeder also caused progenitor cells redistribution between suspension and adhesive fractions by predominantly binding erythroid and multipotential progenitors. Optimum expansion protocol for multipotential progenitors was culturing CD34+-rich cell population with BM-MMSC feeding layer for a week which resulted in the increase the amount of the nucleated cells, CD34+ cells, multi-potential progenitors, and colony-forming units by 56, 36, 45, and 60-fold respectively.
Genes & Cells. 2012;7(1):40-48
pages 40-48 views

Personalized cell-based therapy in ophthalmology: the cytokine profile of autologous cell products

Avetisov S.J., Subbot A.M., Antohin A.I., Kasparova E.A., Kasparov A.A., Pavljuk A.S.

Abstract

The content of the main pro- and anti- inflammatory cytokines and growth factors in autologous cell products, designed for personalized cell-based therapy in corneal endothelium lesions was studied. It was established that cell product contains substances that mediate the effectiveness of cell therapy. The influence of processing conditions on the concentration of cytokines and growth factors in cell products is clarified. It is established that cytokine profile of the cell product is composed of pre-existing substances in freshly blood serum and plasma (TGF-β1, TGF-β2, b-FGF, PDGF-AB, VEGF, IL-4, IF-γ) and those concentration of which increased as a result of processing the cell product - incubation for 4 hours at 37°C (IL-8) and stimulation with polyA:U (IL-1, 6, IF-α and TNF-α). The evaluation of cytokine profile of cell preparation reveals implementation mechanisms of therapeutic effect of personalized cell-based therapy.
Genes & Cells. 2012;7(1):49-53
pages 49-53 views

Cryopreservation human placental tissue as source of hematopoietic and mesenchymal stem cells

Shablii V.A., Kuchma M.D., Kyryk V.M., Onishchenko A.N., Lukash L.L., Lobitseva G.S.

Abstract

In this paper we have show the possibility of cryopreservation of placental tissue for the reason to obtain viable hematopoietic progenitor cells and multipotent mesenchymal stromal cells. It is established, that the relative content of the cells of CD34+/ SSClow/CD45low cells significantly higher in the cryopreserved placental tissue. Also we first described the differences expression of CD90 and CD31on the CD34+/CD45low/SSClow cells of umbilical cord blood and placenta. Clonal analysis of cells revealed the presence in cryopreserved placental tissues of precursors of granulocytes, monocytes and erythrocytes that forming colonies of granulocyte-monocyte, monocytes, granulocytes and erythrocytes. Also it was obtained cultures of stromal cells from cryopreserved placental tissues with immunophenotype CD90+/CD73+/CD105+/HLA-ABClow/CD45-/ CD34-/CD133-/CD14- and adipogenic and osteogenic potential in vitro.
Genes & Cells. 2012;7(1):54-62
pages 54-62 views

Regeneration of rat cardiac myocytes in vitro: colonies of сontracting neonatal cardiomyocytes

Golovanova T.A., Belostotskaya G.B.

Abstract

In parallel to the hypertrophy of major cardiac myocyte population, we detected immunohistochemically the formation of colonies consisting of small (dm = 6,20,5 ƒm) resident c-kit+ and Sca+ stem cells (SC) and Isl1+-positive cardiac myocyte progenitors (CMP) in the primary culture of neonatal rat cardiac myocytes. First contracting colonies (~1-2 clones per 100000 cells) were registered starting from 8th day of culture. The cells of the colonies were capable of spontaneous differentiation, demonstrating the maturation of contractile machinery and Ca2+ responses caffeine (5 мМ) and K+ (120 мМ). The full-scale development of electromechanical coupling with typical for cardiac muscle Ca2+-induced Ca2+ release was obvious at 3 weeks of culture. At first, the local, weak, spontaneous, asynchronous, and arrhythmic contractions at a rate of 2-3 beats/min were registered. However, with time the contractions became synchronous and involved all cells of the colony with the rate of contractions being 58-60 beats/min at the end of the month. First contracting clones comprised Isl1+ CMP, while c-kit+-colonies started to contract 9-10 days later possibly owing to a more prolonged period of proliferation. Thus, we first demonstrated and characterized the contracting colonies originating from SC and CMP when those were co-cultivated with mature cardiac myocytes. The process described in this study is akin to regenerative cardiomyogenesis encompassing the pathway from resident progenitor cell to the colony of mature contracting cardiac myocytes. It follows, therefore, that contracting myocyte colony is a suitable model for basic research, testing of drugs, and the investigation of regenerative capacity of SC and CMP aimed at future applications of resident progenitor cells in cell-based treatment of cardiac injury.
Genes & Cells. 2012;7(1):67-72
pages 67-72 views

An in vivo study of pha matrices of different chemical composition: tissue reaction and biodegradation

Shishatskaya E.I., Nikolaeva E.D., Goreva A.V., Brigham C.J., Volova T.G., Sinskey A.J.

Abstract

The study addresses consequences of subcutaneous implantation of film matrices prepared from different PHAs to laboratory animals. No negative effects of subcutaneous implantation of PHA matrices on physiological and biochemical characteristics of the animals were determined. Independently of the matrices composition and duration of the contact with the internal environment of the organism we did not observe any deviations in the behavior of animals, their growth and development, as well as blood functions. Response of the tissues to PHA matrices was comparable with the response to polylactide, but substantially less expressed at the earlier time periods after implantation. Tissues response to implantation of PHA of all types is characterized by short-term (up to 2 weeks) post-traumatic inflammation with formation of fibrous capsules by 30th-60th days with the thickness less than 100 microns, which get thinner down to 40-60 microns by 180th day as the result of involution. No differences in response of tissues and the whole organism were observed for the matrices produced from the homopolymer of 3-hydroxybutyric acid (P3HB), copolymers of 3-hydroxybutyric and 4-hydroxybutyric acids (P3HB/4HB), 3-hydroxybutyric acid and 3-hydroxyvalerianic acids (P3HB/3HV), 3-hydroxybutyric and 3-hydroxyhexanoate acids (P3HB/3HH). Macrophages and foreign-body giant cells actively participate in the response of the tissues to PHAs. In the studied conditions matrices from the copolymers containing 3-hydroxyhexanoate and 4 hydroxybutyrate were determined as more actively degraded PHA. The next less degraded matrices were matrices from the copolymer of P3HB/3HV and the most resistant were P3HB matrices. The slower degradation of PHA matrices was accompanied by delayed development of giantcells response. The studied PHA matrices can be placed in the following range by their degradation: P3HB/3HH - P3HB/4HB - P3HB/HV - P3HB.
Genes & Cells. 2012;7(1):73-80
pages 73-80 views

Comparative experimental and morphological study of biological osteoplastic materials in bone defects repair

Ter-Asaturov G.P., Lekishvili M.V., Bigvava A.T., Adzhiev K.S., Pankratov A.S., Ryabov A.Y., Yurasova Y.B.

Abstract

The article presents a comparative experimental data of morphological studies with biological plastic materials, actively used in various fields of reconstructive surgery in Russia. Area of implantation was the lower jaw of rabbits, made by original equipment of the model and experiment. Observation periods were 10, 20, 30, 60 and 90 days. Based on morphological analysis of the regenerative abilities assessed followed used materials «Osteomatrix», «CollapAn», «Osteoplast-T» and «Perfoost».
Genes & Cells. 2012;7(1):81-85
pages 81-85 views

Application of electroanalytical techniques for assessment of human cells

Saifullina D.V., Shakhmaeva I.I., Bondar O.V., Abdullin T.I.

Abstract

Rapid development of cell technologies stipulates the need for informative and express methods for the analysis of viability and physiological activity of human cells. We studied analytical possibilities of novel tools for comprehensive characterization of bioelectrochemical properties of living cells, e.g. surface charge of cellular membrane and redox activity of metabolites. Using Malvern Zetasizer dynamic light scattering analyzer we proposed an approach to assessment of zeta potential of human cells and detection of phosphatidylserine on their surface as an early apoptotic marker. On the basis of modified electrodes we designed sensors exhibiting high sensitivity towards electroactive cellular metabolites including antioxidants and macroergic molecules. The sensors were applied for assessment of metabolic activity/energetic status of human cells (blood cells and cell cultures). Electrochemical signal of adenine nucleotides of cells on sensor surface correlated with intracellular level of ATP according to luciferase assay and was found to be more sensitive to alteration in cell viability than conventional MTS test. On the basis of disposable screenprinted electrodes we fabricated a prototype of portable analyzer for rapid analysis of cell health (e.g. for donor cells) in less than 5 ƒl volume of cell sample. Proposed tools and methods are of interest in cell transplantology, basic research and cell-based medical diagnostics.
Genes & Cells. 2012;7(1):86-91
pages 86-91 views

Bio-photon mechanism of the activation of cellular programmes: a haemopoietic precursors proliferation

Vladimirsky E.B., Milman V.D.

Abstract

In this paper the bio-photonic mechanism of the activation of cellular programmes is analyzed. Our basic hypothesis is that cells and individual molecules radiate certain waves that are close to monochromatic (i.e., have a narrow range of radiation frequencies). The cell receptors, for their part, take in certain monochromatic signals. In order to estimate the value of the intensity of radiation it is necessary to refer to the fundamental laws of electro-magnetic waves propagation and photon emission. The research was conducted using a model of the haemopoietic precursors proliferation induction and the apoptosis of the mononuclear cells of peripheral blood ex vivo. The granulocytic colony-stimulating factor (G-CSF) in different concentrations, including ultra-low (homeopathic) ones, was used as the inductor of the proliferation and the inhibitor of apoptosis. Some homeopathic preparations in high dilutions were also included in the study. The study showed the high efficacy of low and ultra-low (homeopathic) concentrations of the preparations. This phenomenon may be explained with the help of the destructive and constructive interference theory. A calculation of the interaction of the monochromic waves radiated by active molecules was done based on their size and molecular mass as well as the size of the target cells. The authors developed an original method of calculating the effective concentration of the preparation.
Genes & Cells. 2012;7(1):92-96
pages 92-96 views

Clinical and experimental study on the use of combined cell transplant on the basis of multipotent mesenchymalstromal cells of adipose tissue in patients with severe deficiency of jaws bone tissue

Alexeeva I.S., Volkov A.V., Kulakov A.A., Goldshtein D.V.

Abstract

The clinical and experimental study on the restoration of bone tissue with tissue engineering scaffold based on stem cells of adipose tissue and resorbable osteoplastic matrix. In a pilot study proved the effectiveness of the transplant, founded the dates of dental implantation. Use of tissue engineering scaffold led to organotypic reconstruction of bone tissue in the field of transplantation in patients with severe atrophy of the alveolar process of the maxilla and mandible. As part of a clinical trial were treated 20 patients.
Genes & Cells. 2012;7(1):97-105
pages 97-105 views

From organ transplantation to reparative spheroids and «microtissues» in suspension 3D-culture

Repin V.S., Saburina I.N.

Abstract

4 experimental alternatives has been developed for organ transplantation: 1. Cell therapy as a suspension of disorganized somatic and stem cells. 2. The Grafts of well differentiated cells on 3D-matrix to compensate (restore) the damaged function. 3. Translational medicine - single selected pilot project of personalized medicine of auto- stem cell sphere transplantation with a new emergent potential for tissue morphogenesis and regeneration. 4. High throughput cellular serial fabrics for multiple generation of standard micro-tissues and 3D-mini-tissues. They are especially devoted to this essay.
Genes & Cells. 2012;7(1):106-108
pages 106-108 views

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