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Vol 8, No 2 (2013)


Prospects for the use of pluripotent stem cells derived blood components: erythropoiesis

Philonenko E.S., Lagarkova M.A., Kiselev S.L.


We review the problem of producing erythroid cells from human pluripotent stem cells. Pluripotent stem cells (embryonic stem cells, and induced pluripotent stem cells) are capable of self-renewal and differentiation into all cell types of an organism, including blood cells. Since a worldwide problem of blood donation shortage is very serious, the development of technologies to produce blood from pluripotent cells is a very modern and necessary task, which is the subject of investigations of many laboratories around the world.
Genes & Cells. 2013;8(2):6-12
pages 6-12 views

Stem cells in carcinogenesis of glioblastoma multiforme

Bryukhovetskyi I.S., Bryukhovetskyia A.S., Kumeiko V.V., Mischenko P.V., Khotimchenko Y.S.


Glioblastoma multiforme is a malignant primary brain tumor with a very poor prognosis. Neural stem and progenitor cells of the adult brain, as well as other types of stem cells are considered by carcinogenesis researchers the most likely source of malignant gliomas. This is evidenced by common genes regulating key processes of life, uniform proteomic profiles and identical immunophenotypical cell surface markers. These cells are highly proliferative, multipotent, able to independently migrate to the damaged area and have extensive replicative potential. However, antitumor properties of the stem cells (SCs) are also confirmed. The bodies of adult mammals and humans have genetically fixed mechanisms of control over their populations and multiple levels of antitumor protection. So far, the role of autologous SCs in a tumor patient is not clear. On the one hand, they fail to fulfill their anti-tumor and regulatory functions, and instead of organizing anti-tumor response become one of the key elements of carcinogenesis contributing to the development of neoplastic tumor vascular network and modulating the processes of neurogenesis, which is the main source of its pathological reinnervation, and therefore, a pain. On the other hand, autologous stem cells trigger powerful proliferative processes in the tumor tissue becoming the driving force of neoplastic growth. Apparently, due to the interaction of tumor cells with autologous stem cells, general and local interstitial patterns of autoregulation and sanogenesis are disturbed making tumor growth possible in principle. From this perspective, the technology that directly affects the population of cancer stem cells seems the most promising.
Genes & Cells. 2013;8(2):13-19
pages 13-19 views

Application of cell and tissue cultures for potential anti-cancer/oncology drugs screening in vitro

Mingaleeva R.N., Solovieva V.V., Blatt N.L., Rizvanov A.A.


Ne of the reasons for the failure of potential anticancer drugs in clinical trials is the imperfection of existing preclinical screening systems. Perhaps the most important step is in vitro testing during which several substances with certain properties should be selected from a large number of substances. An effective system of screening should closely resemble the organization of naturally occurring tumors. Cell cultures are the most simple from technical point of view in vitro models of tumors. However, in many respects cell cultures different from natural tumors. Several models which are more accurately (compared to simple monolayer cultures) emulate the tumor and its microenvironment are developed. An example is three-dimensional cultures. Furthermore, additional methods of anticancer drugs testing are developed based on tissue slice cultures. This review describes current in vitro models which can be used to test the activity of potential drugs for use in treating of oncological diseases.
Genes & Cells. 2013;8(2):20-28
pages 20-28 views

Cryopreserved multipotent mesenchymal stromal cells stimulate reparative chondrogenesis in degenerated intervertebral disc

Iukhta M.S., Volkova N.A., Zhulikova E.P., Goncharuk E.I.


The choice object for cell therapy of degenerative changes of the intervertebral discs is a cryopreserved multipotent mesenchymal stromal cells (MMSC). The research aim was to evaluate a therapeutic potential of cryopreserved allogenic marrow-derived MMSC in this pathology. Rats with modeled compressive degenerative damage of the intervertebral disc CcVI-VII were introduced by 0.5x10 B native or cryopreserved as well as PKH-26-labeled cells on a collagen sponge in the defect area. Animals with spontaneous recovery and administration of saline were served as a control. Histomorphometric study and fluorescent microscopy of intervertebral disc sections was performed on the 30th day after treatment. There was extremely low regenerative potential in the control groups of animals. Histologically after cell therapy, there was tendency to repair of cracks, fissures, collagen fiber fragmentations of the fibrous ring, which was more pronounced in the case of the native MMSC culture application. The cryopreserved MMSC administration was accompanied by increasing in the fibrochondrocyte number of the dorsal annulus per unit area of the intervertebral disc CcVI-VII slice at 1.25 times relative to the model values. At the same time the fibrous ring height increased by 12.5±3.3%, and densitometric index of cartilaginous tissue — by 64±5.7% relative to the model values. The luminescence in the red range of the spectrum in the form of drops and conglomerates, which was localized in outer parts of fibrous ring, was detected by fluorescent microscopy of intervertebral disc cryostat sections on the 30 day after introduction of labeled native or cryopreserved MMSC, that indicated the safety and partial migration of transplanted and/ or their daughter cells in the intervertebral disc CcVI-VII. Our data showed a stimulating effect of native and cryopreserved marrow-derived MMSC suspension in degenerative intervertebral disc damages.
Genes & Cells. 2013;8(2):29-34
pages 29-34 views

A comparative study of the influence of skin fibroblasts and bone marrow stromal cells included in collagen gel on gingiva regeneration

Barmasheva A.A., Nikolaenko N.S., Samusenko I.A., Orekhova L.Y., Pinaev G.P.


The aim of this research was to compare the influence of dermal fibroblasts and multipotent mesenchymal stromal cells included in type I collagen gel on oral mucosa regeneration. It was shown that cultured cells included in collagen gel allowed receiving the greater volume of soft tissues in 180 days than a standard technique of recession management. Repaired gingiva had a normal histological morphology.
Genes & Cells. 2013;8(2):35-43
pages 35-43 views

Comparison of different methods for generation of functional human cardiomyocytes

Khudiakov A.A., Kurapeev D.I., Kostareva A.A., Malashicheva A.B.


Generation of human cardiomyocytes in vitro is important for basic science and for regenerative medicine. There are a lot of published protocols describing the cardiomyocytes obtaining with different efficacy. However in practice, a researcher is faced with the fact that published methods have to be adapted to a particular laboratory for specific tasks. The aim of this study was to compare three methods for generation of human cardiomyocytes in vitro: 1) from human adult heart sample by purifying myocardial progenitor cells and their subsequent differentiation into cardiomyocyte direction; 2) from human adipose tissue multipotent mesenchimal stromal cells (AT-MSC); 3) from human dermal fibroblasts using reprogramming technology and subsequent differentiation to cardiomyocytes. We have shown relatively low efficiency of cardiac progenitor cells and AT-MSC capacity to give rise to cardiomyocyte lineage. The effectiveness of cardiomyocytes differentiation from dermal fibroblasts by reprogramming technology was significantly higher. The advantages, disadvantages and perspectives of each method are discussed.
Genes & Cells. 2013;8(2):47-55
pages 47-55 views

The study of migration of the multipotent mesenchymal stromal cells in the body of animal with a tumor

Meleshina A.V., Cherkasov E.I., Sergeeva E.A., Shirmanova M.V., Balalaeva I.V., Kiseleva E.V., Zayganova E.V.


The study of stem and progenitor cells role in carcinogenesis is the subject of intense investigation. At the same time fluorescent methods become more common for the studying of the stem cells migration into recipient organism and their involvement in tumor pathogenesis. In this study the interaction between bone marrow multipotent mesenchymal stromal cells (MMSC) and allogenic tumor (Lewis lung carcinoma) was investigated using the methods of the flow cytometry and a confocal laser scanning microscopy. MMSC were isolated from bone marrow of the males GFP+-transgenic mice C57/Bl6. The tumors were implanted in mice C57/Bl6 by subcutaneous injection of Lewis lung carcinoma tumor cells. The day before that myeloablation was performed by irradiation. GFP+-MMSC and bone marrow cells from the males C57/Bl6 were administrated into mice C57/Bl6 systemically on the following day after irradiation. The animals were divided into two groups: 1 — irradiated mice with tumor inoculation and injection of bone marrow cells including GFP+-MMSC; 2 (control) — unirradiated mice with tumor inoculation but without cells transplantation. The fluorescence level of freshly isolated bone marrow cells from animals of the first and control groups was evaluated by flow cytometry on the 7, 12 and 15 days after GFP+-MMSC transplantation. In these groups of animals we analyzed the distribution of the labeled GFP+-MMSC and their descendants in the organs and tumor tissues by laser scanning microscopy also on the 7, 12 and 15 days after transplantation. It was established that in systemic administration donor GFP+-MMSC were absent in the bone marrow and tissues of induced tumors, but at the same time they were able to migrate into recipients accumulating in the potential niches (spleen, liver) and proliferate actively.
Genes & Cells. 2013;8(2):56-63
pages 56-63 views

Histogenesis of solid-pseudopapillary tumor of a pancreas

Paklina O.V., Gordienko E.N., Chekmareva I.A.


Solid-pseudopapillary tumor (SPPO) is a rare tumor of a pancreas with unknown histogenesis. In this regards some theories about its development are discussed including genesis from so called cancer stem cells, which can be derivative of embryonic ovaries primordiums. We studied 11 SPPO and 5 ovarian granulosa cell tumors (GCT) adult type by electron microscopy. A comparative ultrastructural study identified similar features in GCT and SPPO: the presence of two cells types, one of which with characteristic nuclei with deep invaginations nukleolemmy as longitudinal furrows, intercellular contacts in the form of desmosomes, in the tissues quite large number of capillaries in the active state were detected. Among all ovarian tumors developing from cells of sex cord stroma SPPO is the most close to GCT. They are related not only due to morphological similarity, but the biological potential, the clinical course and disease prognosis. Our studies confirm the theory of embryonic migration of ovaries primordiums cells with their subsequent contribution to SPPO histogenesis.
Genes & Cells. 2013;8(2):64-68
pages 64-68 views

Algorithm activity of the Stem Cells Bank-Register in Moscow

Kobzeva I.V., Astrelina T.A., Yakovleva M.V., Karpova E.E., Kruglova Y.A., Lebedeva L.L., Puhlikova T.V., Chumak A.A., Stavcev D.S., Gomzyakov A.E., Boyakova E.V.


Progress in transplantation of allogeneic hematopoietic stem cells (HSC) of umbilical cord blood is associated with development of cord blood banks established to provide for the systematic collection, testing, processing, storage, organization and delivery of samples matching cord blood for transplantation. The aim of the study was to develop the algorithm of the Moscow bank's register of unrelated donor of umbilical cord blood HSC. The article described the biological characteristics of all the samples of umbilical cord blood, entered in the register remunerated donors of HSC of Moscow umbilical cord blood stem cell bank (as of 1cryopreservedcord blood sample). It is shown that a small number of unrelated donor of umbilical cord blood HSC in the register allow to select a compatible (6/6), donor-recipient pair with a frequency of 1:151probability. The proposed algorithm of the Moscow bank-register is recommended to use in creating, upgrading and operation of other bank-registers gratuitous unrelated donor of umbilical cord blood HSC.
Genes & Cells. 2013;8(2):69-74
pages 69-74 views

Experience of Samara hematopoetic cells donor registry

Tyumina O.V., Klyuchnikov D.Y., Volchkov S.E., Trusova L.M.


Transplantation of allogenic hematopoietic stem cells used for treatment of oncological and immune diseases during the late 1970s. Despite of that the matched donor search is the main problem in this field as before. To resolve this problem the bone marrow donor registries were established around the world. In 2010 Bone marrow donor registry (BMDR) was established in Clinical Centre of Cell Technologies. BMDR includes adult donors from many regions of Russia and cord blood units (CBUs) from Samara cord blood bank. At the present day, there are more than 5000 CBUs and 5000 donors. During the work of the registry 44 CBUs were handed to Russian and international transplant centers. In according with received results, the survival rate was about 65%. In relation with increase of oncohematological diseases in Russia the development of national bone marrow donor registry is extremely important.
Genes & Cells. 2013;8(2):75-78
pages 75-78 views

The clinical application of autologous mesenchimal stem cells isolated from fat tissue for the treatment of patients with trophic ulcers of lower extremities

Baranov E.V., Tretyak S.I., Vasilevich I.B., Lobanok E.S., Volotovski I.D.


The clinical data about treatment of patients with low intensity trophic ulcer of lower extremities by local transplantation of autologous multipotent mesenchimal stromal cells isolated from fat tissue were presented. The treatment has resulted in complete repair of wound defects, skin recovery. The approaches on fat tissue sampling, isolation, cultivation, expansion and phenotyping of cells and also the methods of cell transplantation into ulcer defects (cell amount, periodicity of transplantation, and mode of insertion) were elaborated.
Genes & Cells. 2013;8(2):79-84
pages 79-84 views

Comparative proteome mapping of tumor stem cells isolated from U87 glioblastoma, neural stem and multipotent mesenchymal stromal cells of a human: from cataloguing of cell proteins to novel paradigm of proteome-based cell therapy of tumors

Bryukhovetskiy A.S., Shevchenko V.E., Chekhonin V.P., Bryukhovetskiy I.S., Kovalev S.V., Baklaushev V.P., Davydov M.I.


We performed proteome mapping, cataloguing and bioinformation analysis of protein lysates of human neural (CD133+) stem cells (NSC) isolated from olfactory sheath of a nose, multipotent mesenchymal (CD29+, CD44+, CD73+, CD90+, СD34 -) stromal cells (MMSC) isolated from human bone marrow and tumor (CD133+) stem cells (TSC) isolated from the human U87 glioblastoma cell line. We identified 1664 proteins in the examined lysates of stem cells (SC), 1052 (63.2%) of which are identical in NSC and TSC and 607 proteins (36.47%) are identical in MMSC and TSC. Other proteins in U87 glioblastoma TSC are oncospecific or carcinogenesis associated. The biological processes, molecular functions, cell localization and protein signal pathways of the proteins available in all three proteomes were annotated by PubMed, PANTHER, Gene Ontology and KEGG databases. It was shown that gliomaspheres of U87 glioblastoma had only 10 intracellular pathways of signal transduction (IPST) that were not modified by neoplastic process, but only two of them (integrin and focal adhesion pathways) were accessible for regulatory action on gene candidates in TSC nucleus. Carcinogenesis free membrane proteins, intracellular pathways of signal transduction and genes expressing proteins of these pathways in U87 glioblastoma TSC can be viewed as main targets for regulatory effect on TSC. We offer a novel concept of proteome-based complex therapy of tumors.
Genes & Cells. 2013;8(2):85-92
pages 85-92 views

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