ROLE OF IMMOBILIZED EXTRACELLULAR MATRIX PROTEINS IN THE KARYOTYPIC VARIABILITY OF "MARKERLESS" CELL LINES Poljanskaya G.G. Institute of Cytology RAS, St. Petersburg, Russia; poljansk@mail.cytspb.rssi.ru      The essential influence of immobilized extracellular matrix proteins - laminin and fibronectin, on the character of the karyotypic variability of different origin (epithelial-like, fibroblast-like) "markerless" cell lines has been shown. The character of numerical karyotypic variability doesn't depend and the character of structural karyotypic variability depends on origin line has been determined. The concrete karyotypic structure corrects the character of the karyotypic variability, providing the way of adaptation of cell population to the definite substrate. These results essentially extend an idea on importance of immobilized extracellular matrix proteins for cell functions. As the alterations of karyotypic structure critically connect with gene expression, which resulted in changes of different cell prorerties.      Key words: karyotypic variability, cell line, extracellular matrix proteins, laminin, fibronectin, structural variant of karyotype, chromosomal aberrations ESTABLISHMENT, CHARACTERISATION AND USE OF THE CONTINUOUS CELL LINE ICO FROM IMMATURE CARP, CYPRINUS CARPIO, OVARIES T.I. Shchelkunova & I.S. Shchelkunov. All Russia Research Institute for Veterinary Virology and Microbiology, Pokrov, Vladimir Province, Russia; shchelkun@yahoo.com      Data on establishment of the ICO cell line from carp, its morphological and caryological characteristics, cultivation parameters, verification of origin, fish virus susceptibility and the use for detection of virus neutralizing antibodies in fish sera are presented.      Key words: continuous cell line, carp, characterization BIOLOGICAL ACTIVITY OF EXTRACELLULAR HISTONES AND PERSPECTIVES OF THEIR APPLICATION IN BIOTECHNOLOGY O. A. Goryukhina,1 G. P. Pinaev 2 1 Biochemical Faculty St.Petersburg State University, 2 Institute of Cytology RAS St.Petersburg, Russia; pushkeen@gmail.com      A review of the literary as well as proper data action of extracellular histones on the functions of different types of cells is considered. Perspectives of the creating the transport systems on the basis of histones for site- specific delivery of therapeutic agents which poor penetration characteristics through cellular membranes and tissue barriers is considered. An opportunity of using histones immobilized on microspheres for modifying the surfaces intended for cultivation of cells was investigated. It was found that histones immobilizied on microspheres facilitated adhesion, proliferation and cell network formation by cellular interactions and cell contacts with several microspheres. The study showed that the substance may be applied for three-dimensional pore matrices designed for tissue-like cell structures in vitro.      Key words: extracellular histones, carrier proteins, microspheres, adhesion, proliferation of cultivated cells ADAPTATION OF MDCK CELLS TO THE GROWTH IN LOW - AND SERUM - FREE MEDIA R. Podchernyaeva,1 G. Danlibaeva,1 N. Mazurkova,2 O. Baklanova,1 V. Chestkov,3 V. Tabakov 3 1 D.I. Ivanovsky's Institute of Virology RAMS, Moscow, 2 GNC of Virology and Biotechnology "Vector", Novosibirsk's region, 3 Medical Genetics Center RAMS, Moscow, Russia; cells@rambler.ru      After five passages in the low-serum rice medium, the adapted to the growth in this medium MDCK cells were obtained. These cells exhibited a decrease in the level of a proliferative activity and morphological changes in comparison with control cultures. Cells preserved viability and activity of reproduction influenza viruses A (H1N1 and H3N2). For the adaptation MDCK cells to the whole serum-free medium "Hybris-2" required 17 passages. At the adapted cells also observed decrease in the level of cell's proliferative activity and changes in morphology of the culture. Reproduction of influenza viruses A (H1N1, H3N2) was lower on 0,5 lg TCD50 in comparison with control, but influenza virus B active reproducted in the adapted and control cells. Therefore a rice medium as well as "Hybris-2" are valid for the cultivation MDCK cells and may be used to the reproduction of influenza viruses.      Key words: cells, adaptation, rice medium, serum-free medium EFFECT OF EXTRACELLULAR MATRIX COMPONENTS ON CLONAL GROWTH AND OSTEOGENIC DIFFERENTIATION OF RAT MESENCHYMAL STROMAL CELLS O.N. Khnykova, O.V. Pajushina, N.N. Butorina, E.I. Buejeverova, V.I. Starostin Koltzov Institute of Developmental Biology RAS, Moscow, Russia; payushina@mail.ru      Proteins of extracellular matrix are essential components of microenvironment of mesenchymal stromal cells (MSC). Investigation of adhesive interactions of MSC with their matrix is crucial for elucidating the mechanisms that regulate proliferation and differentiation of MSC; it is also important for development of cells therapy. The aim of this study consisted in analyzing the influence of some matrix proteins (collagen I, laminin, fibronectin) on clonal growth and osteogenic potencies of rat MSC derived from adult bone marrow or fetal liver. It was found for both bone marrow and fetal liver cells that proportion of fibroblast colony-forming units (CFU-F) adhered for the first 7 days of incubation was significantly higher on fibronectin or collagen I in comparison with uncoated plastic. Laminin affected the adhesion of CFU-F to a lesser degree. Estimation of the alkaline phosphatase activity characterizing early stages of osteogenesis demonstrated decreased osteogenic potential of the bone marrow CFU-F adhered to fibronectin as compared with plastic, collagen I or laminin. The proportion of osteogenic cells among fetal liver CFU-F was extremely low and did not depend on the presence of matrix proteins. Analysis of terminal differentiation of bone marrow MSC in osteogenic medium revealed inhibitory effect of fibronectin on calcium deposition in the foci of osteogenesis. Neither collagen I nor laminin exerted significant influence on osteogenic differentiation in this experimental system. Researching molecular mechanisms of the revealed effects is a promising subject for subsequent experiments.      Key words: mesenchymal stromal cells, colony-forming units, osteogenesis, adhesive interactions, extracellular matrix proteins, collagen I, laminin, fibronectin HLA-ANTIGEN CHARACTERIZATION OF NATIVE AND CULTIVATED NEUROCELLS OF FOETAL AND POSTNATAL HUMAN BRAIN Lyubych L.D., Lisyany N.I., Semenova V.M., Stayno L.P. Acad. A.P. Romodanov Institute of Neurosurgery Acad.Med.Sci. of Ukraine, Kyiv, Ukraine; liubichld@mail.ru      The purpose of the work was study of HLA expression by human neurocells (NC) of different hestation term and differentiation on the protein level and mRNA level using methods of immunophenotyping and RT-PCR. HLA antigens class І, ІІ provide the antigen presentation and recognition thus immunocompetent cells ability to recognize transplanted neurocells, specific response induction and next survival or resection depend on HLA expression by transplanted neurocells. HLA-A,B,C+ and HLA-DR+-expressing cells rate was the lowest among human neurocells of 5 week of hestation and gradually increased to 8-9 week of hestation. mRNA expression of studied allels HLA-A1 and especially HLA-DRa1 increased from full lack or slight amount in human neurocells of 5-9 week of hestation (NSC and neural progenitors) till partially expression in regional NSC of adult brain (Bulbus olfactorius) and significant expression in cells of white matter and especially grey matter of adult brain. Under cultivation in DMEM + retinol acetate the amount and percentage of HLA I and II expressing cells on the protein as well as mRNA levels were decreased whereas the presence in the cultivation medium of the growth factors promoted preservation (FGF) or increasing (EGF) of positive expressing cells amount. Under cultivation in DMEM the HLA I and II expressing Bulbus olfactorius cells number decreased from day 10th to 20th.      Key words: neurocells, bulbus olfactorius, white and grey matter of brain, HLA antigens, immunophenotyping, mRNA expression PRODUCTION OF IgM AND J CHAIN BY B-LYMPHOBLASTOID CELL LINES M. P. Samoylovich, A .A. Pinevich, N. L. Vartanjan, V. B. Klimovich Russian Research Center for Radiology and Surgical Technologies, St.-Petersburg, Russia; mpsamoylovich@gmail.com      One of the main features of B-lymphoblasoid cell lines is production of membrane-associated or secreted immunoglobulins (Ig). Many of these lines secrete IgM molecules that usually contain J chain. Functionally J chain participate in polymerization of pentameric IgM molecules. The lack of J chain or its low expression in the cells results in production of hexameric IgM molecules. Hexamers in contrast to the pentamers can't bind pIgR of epithelial cells and can't form secretory IgM. Detection of J chain in the cytoplasm or in the culture media allows to discriminate between two types of IgM polymers produced by the cells. The aim of the study was the investigation of IgM and J chain production by B-lymphoblastoid cell lines Namalva, NC-37, RPMI 1788, and Raji from Russian collection of cell cultures of vertebrates. ELISA and flow cytometry methods developed on the basis of monoclonal antibodies (MAb) against mu-chains and J chain were used. Wide variability (diversity) of intracellular IgM content was found in RPMI 1788 cell line. The cells expressing J chain comprise less than 30% of total cell number. Cultivation medium from RPMI 1788 contained large amounts of IgM molecules but most of them were devoid of J chain. Populations of Namalva and NC-37 cells were homogeneous in respect of intracellular IgM and J chain content. Low levels of IgM secretion were the characteristics of these two cell lines. Most of IgM molecules in culture media from Namalva and NC-37 were associated with J chain. Raji cells expressed J chain but didn't secrete IgM into culture media. The data obtained allow to conclude that Namalva and NC-37 produce IgM-pentamers, while RPMI 1788 cell line is highly heterogeneous and produces predominantly IgM hexamers.      Key words: IgM, J chain, Namalva, NC-37, RPMI 1788, Raji Instructions for Authors      The paper should not exceed 10 manuscript pages. The manuscript is to be accompanied by recommendation addressed to the Editorial Board of the Information Bulletin from the institution in which the work has been carried out. The paper should contain a brief resume, with the tittle of the work, names of the authors, institution and city.      The manuscript is to be accompanied by its electron variant: one file in MS Word for Windows. 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