RESUMO
A strain of endothelial cells derived from a single cell cloned from a line of normal adult rat lung parenchyma has been maintained in tissue culture for more than 3 years. These cells have been identified as endothelial cells based on the combination of their growth characteristics, cell morphology as observed with both light and electron microscopy, and their physiological properties. They have continued to produce granules, which stain specifically for glycosaminoglycans with Alcian blue, for over 2 1/2 years. During the same period of time, glycosaminoglycans were identified biochemically in both cells and medium. They have maintained the ability to degrade bradykinin over this period as well.
Assuntos
Células Clonais , Pulmão/citologia , Animais , Bradicinina/metabolismo , Divisão Celular , Linhagem Celular , Células Clonais/enzimologia , Células Clonais/ultraestrutura , Grânulos Citoplasmáticos/ultraestrutura , Endotélio/citologia , Glicosaminoglicanos/análise , Lisina Carboxipeptidase/metabolismo , Ratos , Vacúolos/ultraestruturaRESUMO
Antibodies were developed in rabbits against an established line of endothelial cells from normal adult rat lung. Pre- and postimmunization sera were tested for antibody activity by the indirect immunofluorescence technique. Preimmunization serum failed to react with the endothelial cells, whereas the antibody titer of postimmunization serum from two rabbits was 1:512. Organ specificity and species specificity were assessed by absorbing the serum with packed dissociated cells from different organs of the rat and lung cells of other species. Only cells obtained from rat lung absorbed the antibodies completely. The antiserum showed some crossreactivity with the other cultured cells but the pattern of fluorescence was different. In the presence of complement the antiserum was found to be cytotoxic to cultured rat lung endothelial cells but not to the other crossreacting cells.
Assuntos
Anticorpos , Pulmão/imunologia , Animais , Especificidade de Anticorpos , Antígenos , Membrana Celular/imunologia , Células Cultivadas , Proteínas do Sistema Complemento , Reações Cruzadas , Citotoxicidade Imunológica , Endotélio/imunologia , Epitopos , Masculino , Especificidade de Órgãos , Coelhos , RatosRESUMO
The glycosaminoglycans produced by a mixed cell line of normal adult rat lung and an endothelial clone derived from this line were isolated and examined. Cellulose acetate electrophoresis of media and cells before and after digestion with specific enzymes indicated that all the major glycosaminoglycans except keratan sulfate were synthesized by both cultures. Heparan sulfate and dermatan sulfate were found only in the cell fraction while hyaluronic acid was found in both the medium and the cell fractions. The chondroitin sulfates were isolated from the medium. The endothelial clone produced a 4:1 ratio of glucosamine to galactosamine in the medium from the fifth through thirteenth months of culture. The medium of the mixed cell line initially contained glycosaminoglycans with a glucosamine to galactosamine ratio of 2:1 but after approximately one year of culture, the ratio had changed to 4.6:1 suggesting that the culture contained predominatly endothelial cells.
