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CELL CULTURES
Information Bulletin. Issue 31, St. Petersburg, 2015 |
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COMPARATIVE EVALUATION OF THE RAT NEUROGENIC SUPERNATANT AND PREPARATION GALAVIT
CYTOTOXIC EFFECT ON CULTURES OF HUMAN GLIOBLASTOMA
Treatment of patients with malignant brain gliomas remains one of the unsolved and urgent problems of modern neurooncology. One of the directions, actively developed to optimize the treatment of these patients is the search for drugs that can stimulate anti-tumor immunity and inhibit tumor proliferation.
Key words: supernatant of rat neurogenic cells, galavit, human glioblastomas, cytotoxic index, mitotic index. COMPARATIVE STUDY OF MORPHOFUNCTIONAL ACTIVITY OF THE BONE MARROW CELLS, CULTIVATED
IN VIVO AND IN VITRO, OF PATIENTS WITH CHRONIC MYELOID LEUKEMIA DURING THE TREATMENT WITH TYROSINE KINASE INHIBITORS
In this study we have assessed the morphological and functional activity of bone marrow cells of patients with chronic myeloid leukemia (CML) in semisolid agar in vitro and in vivo. Tyrosine kinase inhibitors of the first- and the second-line were used in the therapy. The study of hematopoietic bone marrow cells were produced through stimulation with in vitro culture with adding the granulocyte-macrophage growth factor to culture medium. Culturing cells in vivo was performed in special diffusion chambers, placed in the peritoneal cavity of experimental animals (mice). In all variants of experiment colonies and clusters were counted in culture and their morphological composition on drugs colored using Pappenheim method was assessed. The maturation index of granulocytes was also counted. These results indicate that the number of colonies in semisolid agar in vitro and in vivo in patients with an optimal response to therapy with imatinib and nilotinib was significantly lower than in patients with acquired drug resistance and in patients with newly diagnosed CML. Furthermore, it was found that in patients, who took TKI therapy as mentioned above, the number of clusters in semisolid agar in vivo was significantly greater than the number of such cells in in vitro culture. This may indicate the influence of the microenvironment factors which are present in the body of the recipient animal on the proliferation of leukemic clone of committed cells with limited proliferative potential. This fact has received ΰ further evidence with a shown difference in the index of maturing of granulocytes in culture systems in vitro and in vivo. Key words: chronic myeloid leukemia, targeted therapy, microenvironment factors, cell culture in semisolid agar in vitro and in vivo ZAJDELA HEPATOMA CELLS IN CULTURE: CLONING AND CHARACTERIZATION OF CLONAL LINES
Rat ascitic Zajdela hepatoma is a metastatic tumor, or, more precisely, the phase of the metastatic cascade, and can be used as a promising model for studying the cellular mechanisms of tumor progression and metastasis. Work was carried out on two cultured in vitro lines of Zajdela hepatoma with different levels of cytodifferentiation and tumorogenesis monolayer (ZH-ad) and suspension (ZH-fl), represented by the floating multicellular Islands. To identify cell subpopulations with tumor initiating potential and/or ability of invasion and metastasis in the cell lines, the cloning of cells was carried out by the method of limiting dilutions. It was shown that both cell lines generate three types of clones which differ in the form of colonies and proliferative potential; it should be noted that for the first time it was revealed the formation of spherical clones in complete growth medium when cloning of cell suspension line. Of the eight received clone lines only the cells of spherical clone and two holoclones induced the development of ascitic tumors after intraperitoneal injection to outbred rats. First established that the cells of the monolayer and suspension lines and their clones differ in the expression pattern of epithelial cell adhesion molecule (EpCAM), which is a marker of liver stem cells and cancer stem cells (CSC), or tumor initiating cells, and the activity of matrix metalloproteinase (MMP) 9, the level of secretion which may serve as an indicator of invasiveness of tumor cells. Identification of tumorogenic spherical clones and holoclones with different adhesive properties, and MMP activity allow to make a conclusion about the heterogeneity of the subpopulation of tumor initiating cells of ascitic hepatoma Zajdela. Key words: hepatoma, the cloning of cells, tumor stem cells, tumor initiating cells, matrix metalloproteinase DETERMINATION OF THE WHEAT EMBRYO AUTONOMY BY THE EMBRYO CULTURE
IN VITRO METHOD
In the experimental conditions of embryo culture in vitro, using the hormone-free nutrition medium, it was showed that the formed wheat embryo (17.520 days after pollination, 2.12.2 mm of length) characterized by availability of all typical cereal embryo organs is in keeping with the stage of autonomy. Key words: embryo culture in vitro, embryo autonomy, spring soft wheat IMMUNOLOCALIZATION OF CYTOKININS IN WHEAT POLYEMBRYOIDS IN VITRO AT THE INITIAL STAGES
OF DEVELOPMENT
Under experimental conditions of in vitro culture the immunolocalization of endogenous cytokinins in cells of morphogenetic centers and meristematic zones of callus, as well as in cells of emerging shoot and root apices of developing polyembryoids of wheat were revealed. These data suggests the involvement of cytokinins in the initial stages of polyembryoidogenesis in vitro in wheat. Key words: culture in vitro, callus, polyembryoid, spring soft wheat CHARACTERIZATION OF THE KARYOTYPE OF DXB-11 CELL LINE, DERIVATIVE OF
CHINESE HAMSTER OVARY CELL LINE CHO-K1
TChinese hamster ovary (CHO) cells are a source of the biotechnological production of a wide range of proteins used for pharmacological purposes. Karyotypic diversity of CHO cell lines, their long-term cultivation in non-standardized culture conditions in different laboratories determines the significance of cytogenetic studies. Here, the karyotype of the dihydrofolate reductase-deficient DXB-11 cell line (derivate of the CHO-K1 cell line) from the Russian cell culture collection is described for the first time. Cytogenetic analysis has been performed by G-banding. Cells of the modal class (78%) had 20 chromosomes, and were characterized by karyotypic heterogeneity caused by non-clonal chromosome rearrangements. Fourteen abnormal (including 8 unidentified, marker chromosomes) and only 6 normal chromosomes were revealed in the DXB-11 karyotype. G-banding patterns of DXB-11 and parental CHO-K1 (Filatov, Mamaeva, 1985) cell lines have been compared. Chromosomes 1, 5 in two copies both, and also chromosomes 8, 9, and abnormal chromosomes del(2)(q11q22), add(6), der(X) and 5 marker chromosomes were common for DXB-11 and CHO-K1 cell lines. Chromosomes del(2)(p16p25), der(6), mar7 θ mar8 were specific only for DXB-11 cells. Appearance of these new chromosomes in the DXB-11 karyotype resulted from the structural rearrangements of normal chromosomes 2 and 7 and of one abnormal chromosome of CHO-K1 karyotype. As a whole, similarity of the DXB-11 and CHO-K1 karyotypes gives evidence of stability of the genetic material in CHO cells for more than 50 years of maintenance in vitro. Key words: chinese hamster ovary DXB-11 cell line, karyotype, karyotypic heterogeneity INFLUENCE OF ANTIVIRALS ON PROLIFERATION, APOPTOSIS AND DIFFERENTIATION OF CULTURED HUMAN CELL LINES
Cultured cell lines are widely used for experimental testing of antivirals. The present study explored the influence of antivirals ingavirin, triazavirin, ribavirin and rimantadine on proliferation, apoptosis and differentiation of human cell lines. The experiments were done using normal and transformed cell lines: FLECH (embryo human lung diploid fibroblasts), ECV-304 (spontaneously transformed endothelial cells), A-549 (human lung carcinoma) and K-562 (myelogenic human leukemia cell line). Influence of antivirals on differentiation of hemopoetic cells was studied on K-562 cells using hemoglobin as a marker of differentiated state.
Key words: cell lines FLECH, ECV-304, A-549, K-562, antivirals, proliferation, apoptosis, and differentiation THE CHANGE IN GROWTH ACTIVITY AND VIABILITY OF CULTURED HUMAN EMBRYO FIBROBLASTS UNDER
THE INFLUENCE OF POLYOXOMETALLATES
Comparative testing of 4 products POM (polyoxometallates) in cultured human embryo fibroblasts (FEH-T) showed different toxic effects on the cells studied centimolar solutions
H3PW12O40, H4SiW12O40, H4SiMo12O40 and
H3PMo12O40. Using the MTT test showed the reduction in cell proliferation during incubation with the studied POM at a concentration of
160.620 μM/ml. Most cytotoxicity after 24 hours was found in H4PW12O40 containing tungsten, the smallest was in
H4SiMo12O40 with molybdenum, that may indicate the degree of exposure depending on their physicochemical properties.
Key words: nanoparticles, polyoxometallate, cell proliferation, electron microscopy, cytokines |
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