Xeno-miRNet: a comprehensive database and analytics platform to explore xeno-miRNAs and their potential targets

A new method is presented for the quantitative analysis of intercellular adhesive specificity. In this assay, two cell types are mixed, one unlabeled and the other labeled with the fluorescent dye, fluorescamine [4-phenylspiro(feran-2[3H],1'-phthalan)-3,3'-dione]. The resulting aggregates are analyzed by fluorescence microscopy to determine the number of labeled and unlabeled cells per aggregate. Random (nonspecific) aggregation was characterized by a binomial distribution, and adhesive specificity was accordingly quantified by the deviation (as determined by a chi-square test) from the calculated binomial distribution. The labeling procedure was simple and rapid, and experiments with 18 different cell types showed that it did not affect cell viability, morphology, rate and extent of adhesion, plating efficiency, and the capability of myogenic cells to undergo terminal differentiation. Most important, assays with morphologically identifiable cell pairs indicated that the fluorescent label neither induced apparent nor destroyed existing adhesive specificity. The most pronounced adhesive specificities were observed with freshly explanted cells from adult tissues and also with mixtures of simian virus 40- transformed and nontransformed BALB/c 3T3 cells. A glucosamine-6- phosphate N-acetylase-deficient mutant 3T3 line (AD6), however, aggregated randomly with parental 3T3 cells. Lectin-resistant mutant Chinese hamster ovary (CHO) cells displayed marginal adhesive specificity when mixed with normal CHO cells.

[1]  Malcolm S. Steinberg,et al.  Reconstruction of Tissues by Dissociated Cells , 1963 .

[2]  S. Roseman,et al.  A requirement for reversible binding between aggregating embryonic cells before stable adhesion. , 1975, The Journal of biological chemistry.

[3]  C H Wang,et al.  Studies on the mechanism by which cyanine dyes measure membrane potential in red blood cells and phosphatidylcholine vesicles. , 1974, Biochemistry.

[4]  R. Hynes,et al.  Characteristics of concanavalin A‐resistant Chinese hamster ovary cells and certain revertants , 1979, Journal of cellular physiology.

[5]  I. Pastan,et al.  Mutants of mouse fibroblasts altered in the synthesis of cell surface glycoproteins. Preliminary evidence for a defect in the acetylation of glucosamine 6-phosphate. , 1977, The Journal of biological chemistry.

[6]  P. Stanley,et al.  Selection and characterization of eight phenotypically distinct lines of lectin-resistant chinese hamster ovary cells , 1975, Cell.

[7]  H. Eppenberger,et al.  Differentiation in cultures derived from embryonic chicken muscle. II. Phosphorylase histochemistry and fluorescent antibody staining for creatin kinase and aldolase. , 1976, Developmental biology.

[8]  I. Pastan,et al.  Role of cell surface carbohydrates and proteins in cell behavior: studies on the biochemical reversion of an N-acetylglucosamine-deficient fibroblast mutant. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[9]  L. Sjöström,et al.  Isolation and characterization of cells from rat adipose tissue developing into adipocytes. , 1978, Journal of lipid research.

[10]  K. Vosbeck,et al.  Assay of intercellular adhesiveness using cell-coated Sephadex beads as collecting particles. , 1976, Journal of cell science.

[11]  S. Bozzaro,et al.  Contact sites in aggregating cells of Polysphondylium pallidum. , 1978, Journal of molecular biology.

[12]  D. Roncari,et al.  Cytological and enzymological characterization of adult human adipocyte precursors in culture. , 1976, The Journal of clinical investigation.

[13]  M. Bissell,et al.  The use of fluorescamine as a probe for labeling the outer surface of the plasma membrane. , 1976, Biochemical and biophysical research communications.

[14]  S. Roseman,et al.  AN ASSAY FOR INTERCELLULAR ADHESIVE SPECIFICITY , 1971, The Journal of cell biology.

[15]  K Weber,et al.  Cytoplasmic microtubular images in glutaraldehyde-fixed tissue culture cells by electron microscopy and by immunofluorescence microscopy. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[16]  L. Glaser,et al.  Surface components and cell recognition. , 1979, Annual review of biochemistry.

[17]  J. Holtfreter.  Gewebsaffinität, ein Mittel der embryonalen Formbildung , 1939 .

[18]  G. Rothblat,et al.  Release of lipoprotein lipase from rat adipose tissue cells grown in culture. , 1977, Biochemical and biophysical research communications.

[19]  S. Grady,et al.  Intercellular adhesive selectivity. III. Species selectivity of embryonic liver intercellular adhesion , 1976, The Journal of cell biology.

[20]  G. Edelman,et al.  Cell fractionation and arrangement on fibers, beads, and surfaces. , 1971, Proceedings of the National Academy of Sciences of the United States of America.

[21]  L. Culp,et al.  Sorting out of normal and virus-transformed cells in cellular aggregates , 1976, The Journal of cell biology.

[22]  M. Takeichi,et al.  Experimental manipulation of cell surface to affect cellular recognition mechanisms. , 1979, Developmental biology.

[23]  S. Roseman,et al.  Adhesive specificity of juvenile rat and chicken liver cells and membranes. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[24]  I. Pastan,et al.  A mutant fibroblast cell line defective in glycoprotein synthesis due to a deficiency of glucosamine phosphate acetyltransferase. , 1978, Archives of biochemistry and biophysics.

[25]  S. Krag.  A concanavalin A-resistant Chinese hamster ovary cell line is deficient in the synthesis of [3H]glucosyl oligosaccharide-lipid. , 1979, The Journal of biological chemistry.

[26]  P. Seglen.  Preparation of isolated rat liver cells. , 1976, Methods in cell biology.

[27]  Philip L. Townes,et al.  Directed movements and selective adhesion of embryonic amphibian cells , 1955 .

[28]  S. Roth,et al.  Adhesive specificity in normal and transformed mouse fibroblasts. , 1973, Developmental biology.

[29]  I. Pastan,et al.  Mutants of Balb/c 3T3 fibroblasts defective in adhesiveness to substratum: evidence for alteration in cell surface proteins. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[30]  R. Marchase,et al.  Intercellular adhesive specificity. , 1976, Biochimica et biophysica acta.

[31]  Sidney Udenfriend,et al.  Fluorescamine: A Reagent for Assay of Amino Acids, Peptides, Proteins, and Primary Amines in the Picomole Range , 1972, Science.

[32]  S. Roseman.  The synthesis of complex carbohydrates by multiglycosyltransferase systems and their potential function in intercellular adhesion. , 1970, Chemistry and physics of lipids.

[33]  S. Roseman,et al.  A quantitative assay for intercellular adhesion. , 1973, Proceedings of the National Academy of Sciences of the United States of America.