Technical Information Production of BioloT, Inc.			3
Stem cells: their use in fundamental cell biology and medicine POSTNATAL NEUROGENESIS IN HUMAN OLFACTORY BULB. V.M. Semenova, V.V. Medvedev. Acad. A. Romodanov Institute of Neurosurgery, Academy of Medical Sciences of Ukraine, Kyiv, Ukraine.      The authors of the review summarize results of the last decade studies on processes of neurogenesis in the adult central nervous system. The main attention is paid to neurophysiology and morphology of olfactory bulb as a source of accumulation of neural stem cells (NSC) that migrate into this structure from the subventricular zone of lateral ventricules and differentiate there into mature neurons and glial cells. The authors analyze evidence for activation of neurogenesis in postnatal animal brain under certain pathological conditions (forebrain ischemia, seizures). They examine perspectives of NSC utilization in treatment of neurodegenerative and posttraumatic diseases of the central nervous system.			5
CHARACTERISTICS OF THE PRIMARY CULTURE OF BROWN ADIPOCYTES. G.E. Bronnikov. Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Moscow District, gennady_bronnikov@rambler.ru      Brown adipose tissue hyperplasia is a possible way to restore the ability of adult individuals to dissipate energy as heat that may prevent obesity in the case of overeating. Primary culture of brown adipocytes is a good model for investigation of molecular events of the intracellular signaling and a search for a specific medication that could be useful for in vivo stimulation of the brown preadipocyte proliferation. Cultured brown preadipocytes are able to spontaneously proliferate and, on reaching confluence, to differentiate to result in mature brown adipocytes. Both cell proliferation and cell differentiation can be stimulated in cultured cells by norepinephrine via different types and subtypes of adrenergic receptors.			13
KARYOTYPE HETEROGENEITY OF EMBRYONIC STEM CELLS OF THE MOUSE LINE ES R1. Т.Т. Glazko, L.M. Mezhevikina*, А.A. Boiko. Institute of Agroecology and Biotechnology UAAN, Kiev, Ukraine, glazko@biotech.relc.com; * Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Russia, mezhevikina@rambler.ru      Analysis of cytogenetic anomaly frequencies in three sublines of ES R.1 line and its 5th clonal sublines obtained from two cell colonies after transformation of the ES R.1 cells by plasmid with the gene Lif was carried out. Cell transformation did not lead to an increase of cytogenetic anomalies; however, initial sublines of the ES R.1 line, as well as its transformed clonal descendants, had the redundant quantity of the chromosome 8 material in the structure of various Robertsonian translocations even in cells with the diploid chromosome number (2n = 40).      The obtained data allowed us to put forward the following hypothesis about the "chain" karyotype evolution of the ES cells in vitro. The cell division in the cultural medium with cytokines promotes among them selection of cell clones with increased "sensitivity" to cytokine effects. The cells with gain of the chromosome material with such genes are picked up by selection. Since the genes of chromatide segregation control also are located on the same chromosomes, this leads to further disturbance of normal mitosis passing and accumulation of cell clones with a broad spectrum of combinations of excess on any other chromosomes, which, in turn, is a substrate of further selection. Such scheme of passive involvement in selection of genes, linked with loci, which is the target of selection, is well known in population genetics as "hitch-hike" effects. According to our obtained data, similar events can be realized in the karyotype evolution of ES cells.			19
Cell Biotechnology and Tissue Engineering PROLIFERATION AND DIFFERENTIATION OF HUMAN EMBRYONAL AND RAT NEONATAL CARDIOMYOCYTES IN VITRO. I.L. Erokhina, E.G. Semenova, O.I. Emelyanova. Institute of Cytology, Russian Academy of Sciences, St. Petersburg, Russia, heartdev@mail.cytspb.rssi.ru      Cardiomyocytes were isolated enzymatically from the 19-22-week old human embryos and the 2-3-day old rats and cultured for 14 days. DNA synthesis, ultrastructure, and the presence of atrial natriuretic peptide (ANP) were examined. The degree of proliferation and differentiation of human embryonal myocytes is lower than that of neonatal rat myocytes. After a 24-hr incubation with ³H-thymidine in 7-day-old culture, 1.8 ± 0.5% of human embryonal myocytes and 22.0 ± 2.2% of rat neonatal myocytes are labeled. Human and rat cardiomyocytes differentiate, synthesize DNA and ANP within 14 days in vitro; this is indicative of vitality of these cells.			26
CLINICAL USE OF BONE MARROW STEM CELLS IN RECONSTRUCTING MAXILLOFACIAL SURGERY AND DEVELOPMENT OF METHODS FOR THEIR CULTIVATING AND TESTING. A. Bakhtinov, A. Polekhin, E. Lezhnev, A. Shamsoudinov. Stomatology Institute, Moscow; Institute of Theoretical and Experimental Biophisics, Russian Academy of Sciences, Pushchino, Russia, plastikline@inbox.ru      During the experiment a hundred of rats have been defected at the angle of mandible. The defect was filled with a combination of autologous bone marrow and a part of block porous hydroxyapatite obtained as a result of an ox bone burnt at 1100?C. The same combined graft was placed into the muscular pocket on the rat femur posterior surface. The animals, whose defects were filled with hydroxyapatite without the bone marrow, formed the control group. In 3 months, according to results of autopsy, the entire regenerator filling the defect of the mandible was observed. Morphological and biochemical methods prove an advantage of the combined graft over hydroxyapatite.      Since 1999 the autologous bone marrow with hydroxyapatite preparations has been used to fill patients' jaws defects after surgical treatment of keratocysts. The results were followed from a month to 2 years. X-ray examination revealed restoration of bone structures in the defect-filling region in 3-4 month. The main condition for successful development of autotransplantation is to promote cell-growing technologies. The basic ways of all technologies are discussed.			31
Studies on cultured plant cells and tissues WHEAT PLANT REGENERATION IN EMBRYOGENIC CALLUS IN VITRO: ROLE OF EXOGENOUS ABA. E.M. Zainutdinova¹, N.N. Kruglova¹, I.F. Shayakhmetov². ¹Institute of Biology of Ufa Scientific Center, Russian Academy of Sciences, Ufa, Russia, Kruglova@anrb.ru, ²Bashkir State University, Ufa, Russia      The cyto-histological analysis of developing embryogenic calli obtained in wheat immature embryo culture in vitro was carried out. It was shown that morphogenetic center consisted mainly of undifferentiated meristematic cells capable for further development. The amount of morphogenetic centers in embryogenic callus depends both on exogenous ABA concentration and on duration of culturing in vitro. The maximum amount of the morphogenetic centers in embryogenic calli is found at use of exogenous ABA at a concentration of 2.0 mg/L after culturing in vitro for 20 days.			35
Development of Problem METHOD OF DECONTAMINATION OF CELL CULTURES FROM MYCOPLASMAS WITH THE HELP OF FLUOROQUINOLONES IN COMBINATION WITH ANTIBIOTICS. G.R. Mikhailova, T.M. Khizhnyakova, R.Ya. Podcherniayeva, O.M. Grinkevich, T.I. Aliper. D.I. Ivanovsky Institute of Virology, Russian Academy of Medical Sciences, Moscow, Russia, lena_firs@inbox.ru      It is established that fluoroquinolones enrosept and abaktal actively reduce the amount of mycoplasmas in the cells of HeLa and FrhK-4/R from maximal initial concentration to minimal amount or to its complete elimination. These results reveal three subcultivations of the cells with addition of non-toxic fluoroquinolone amounts. At the time of further cultivation of decontaminated cells the maintenance of mycoplasmas did not increase. A combination of fluoroquinolones with antibiotics allows simplification of the methods of decontamination and shortens time of cell treatment.			41
THE DETECTION OF CELL AND INFECTIOUS PRIONS (PRPC-SC/C) IN CELL CULTURES AND BIOLOGICAL FLUIDS. A.S. Simonova, N.B. Savenko, L.P. Dyakonov, V.M. Kolyshkin, A.E. Moshkov, O.M. Volpina. Kovalenko All-Russia Institute of Experimental Veterinary Medicine, Moscow, Russia, levdyakonov@mail.ru      The article reviews data from special literature on methods of detection of prions in tissues, cell cultures, and biological fluids (blood, urine, etc). It also presents results of our own study on immunocytochemical detection of prions in cell cultures and immunofluorescent detection of prions in animal blood serum and protein substrate.			45
Collections of cell cultures ADDITIONAL INFORMATION ABOUT CELL LINES IN THE COLLECTION OF D.I. IVANOVSKY STATE RESEARCH INSTITUTE OF VIRUSOLOGY OF THE RUSSIAN ACADEMY OF MEDICAL SCIENCES. R.Ya. Podchernyaeva, T.M. Khizhnyakova, G.R. Mikhailova, A.D. Petrachev. Ivanovsky Institute of Virology, Russian Academy of Medical Sciences, Moscow, Russia, lena_firs@inbox.ru			51
Methods of investigations FISH CELL CULTURES FOR VIROLOGICAL INVESTIGATIONS AND DIAGNOSIS OF VIRAL DISEASES. E. Zavyalova, T. Pichugina, L. Dyakonov, M. Borisova. Kovalenko All-Russia Institute of Experimental Veterinary Medicine, Moscow, Russia, levdyakonov@mail.ru      Review of the 40-year history of fish cell cultivation in vitro is presented.      Properties of foreign cell lines and primary cell culture and the growth of different viruses in these models are described.			53
GLUCURAL AS A CYTOPROTECTOR. STUDY ON ISOLATED RAT HEPATOCYTES. G. Sakuta¹, E. Baydiuk¹, A. Litanyuk¹, V. Morozov¹, A. Oksman². ¹Institute of Cytology, Russian Academy of Sciences, St. Petersburg, Russia, cellpath@mail.cytspb.rssi.ru, ²Mount Sinai School of Medicine, New York, USA      Isolated hepatocytes are a widely spread object both in fundamental investigations and in practical medicine. The important problem is a search for ways to prolong the hepatocyte life span. The goal of this work was to study effectiveness of Glucural (a water-soluble N-methyl-d-glucosamine complex of 6-methyluracil) as a cytoprotector for isolated rat hepatocytes. Hepatocytes were isolated by a modified two-step in situ collagenase type IV perfusion method. It was shown that Glucural (50-100 µg/ml) had obvious positive effect as hepatocyte cytoprotector when storing hepatocytes. Glucural averts hepatocyte agglutination in suspension. At the same time, this Glucural property decreases hepatocyte spreading on plastic. Thus, Glucural can be used as a cytoprotector for isolated rat hepatocytes when storing them in suspension.			58
Chronicle This section contains information on results of conferences held in 2004 and on conferences planned for 2005. NEW FRONTIES IN FUNDAMENTAL RESEARCH AND CREATION OF CELL TECHNOLOGIES BASED ON HUMAN EMBRYONAL STEM CELLS. O.F. Gordeeva. Koltzov Institute of Developmental Biology, Russian Academy of Sciences, Moscow, Russia, olgagordeeva@yandex.ru			63
Information of the Cell Culture Association			75
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 title of the work, names of the authors, institution, city.      The manuscript is to be accompanied by its electron variant: one file in MS Word for Windows. The file should have the font Arial Narrow, 13; interline difference - 1.5; the indented line - 0.5 cm; the upper margin - 2.5 cm, other margins - 2.1 cm each. The file is to be e-mailed to the address of the Editorial Board.			76