A monoclonal antibody to a cell surface determinant in human endometrial epithelium: stage-specific expression in the menstrual cycle.

A monoclonal antibody (CC25) was obtained after immunization of mice with intact glandular epithelial cells from secretory phase endometrium. Here we report a preliminary immunohistologic study in the endometrium of 29 patients in different phases of the menstrual cycle and early pregnancy. In immunofluorescence, CC25 binds to a basolaterally oriented epithelial cell surface antigen that is absent during the proliferative phase and appears suddenly in both glandular and uterine surface locations soon after ovulation.

Importance of vitamin C in maintenance of the normal amnion: an experimental study.

Collagenous matrix in amnion accounts for most of the dry weight of the tissue and provides its mechanical strength and resistance to rupture. Cell and organ culture techniques have been utilized to study the influence of vitamin C upon the synthesis and deposition of extracellular matrix by cells of normal amnion at term. The cultures have been examined using light and electron microscopy and metabolic labelling.

Monensin-dependent and -independent mechanisms of cell-matrix adhesion.

Attachment and spreading of human FL cells on a subcellular matrix (SCM) preparation made by treating confluent cell monolayers with deoxycholate are insensitive to the presence of monensin. However, if the cell suspension is surface-iodinated prior to adhesion using the LPO/H2O2 system, cell spreading on SCM is inhibited by 1 microM monensin. The suggested interpretation is that cell surface components required for cell spreading on SCM are inactivated by iodination and need replacement from intracellular reserves by a monensin-sensitive pathway.

The extracellular matrix of human amniotic epithelium: ultrastructure, composition and deposition.

Ultrastructural comparisons have been made between human amnion extracellular matrix in tissue and cell culture. Immunochemical analysis of matrix deposited by monolayers of cultured amnion epithelial cells has also been undertaken. The basal cell surfaces are highly invaginated with an associated basal lamina that is more electron dense at the distal tips of basal cell processes where hemidesmosomes are frequent.

An immunofluorescence study of extracellular matrix associated with cytotrophoblast of the chorion laeve.

Using immunofluorescence we have studied the distribution within and beneath the cytotrophoblast of chorion laeve of five extracellular matrix components: types I, III and IV collagen, fibronectin and laminin. Fibronectin, laminin and types I and IV collagen are located in a network of extracellular matrix which encapsulates cells of the cytotrophoblast multilayer. The results suggest that cytotrophoblast in all layers is active in matrix biosynthesis.

Basally located epithelial cell surface component identified by a novel monoclonal antibody technique.

Tubular aggregates of glandular epithelial cells (gland fragments) were isolated from human endometrium by collagenase digestion of surrounding stroma, thus exposing the basal surfaces of the cells. Using these aggregates as immunogen, monoclonal antibodies could be derived that recognized basally located antigens. One such antibody, G71, is described, that binds to a basal epithelial cell antigen present in a variety of human epithelia.

Defective adhesion to extracellular matrix leads to altered social behaviour in cultured fibroblasts.

We describe the properties of variant mouse fibroblasts selected for poor adhesion to growth substratum containing subcellular matrix accumulated by adherent cells at confluence. The variant cells adhere to virgin plastic and grow normally to confluence in the presence of serum. After subculture and reseeding onto the same surface the cells initially adhere, but after a further 2 days of growth they retract into aggregates and detach.

Adhesion of human amnion epithelial cells to extracellular matrix. Evidence for multiple mechanisms.

Human amnion epithelial cells attach and flatten slowly (approximately 65 min) onto plastic in the presence of serum but much more rapidly (20-30 min) onto subcellular matrix (SCM) deposited by the same cells. This matrix contains both fibronectin and laminin, but neither molecule on its own can reproduce its adhesive properties. Cells attach on surfaces containing fibronectin and laminin and extend filopodial and lamellipodial areas of cytoplasm without extensive flattening in the perinuclear region.

Novel substratum-dependent dendritic morphology and motility in epithelial cells.

Qualitative and quantitative light microscopy including time-lapse observations, and scanning electron microscopy are applied to study the response of cultured primary amnion epithelial cells to surfaces coated with a fibronectin-containing aggregate extracted from placenta (PSF). Cell shapes are compared with those observed in medium containing serum and amniotic fluid. Cells respond rapidly to PSF in serum-free medium to give unusual dendritic structures.

Kinetic analysis of cell spreading. II. Substratum adhesion requirements of amniotic epithelial (FL) cells.

The kinetics of spreading of trypsinized FL cells on plastic or glass substrata covalently or passively coated with various proteins to make simplified model extracellular matrices have been measured. Kinetics have also been obtained in the presence and absence of serum and amniotic fluid. Data from such experiments are shown to be sigmoid and have been computer-fitted with great accuracy to 12 mathematical models discussed in the accompanying paper.

Kinetic analysis of cell spreading. I. Theory and modelling of curves.

The spreading of cells on suitably treated surfaces is briefly discussed and the need to analyse this phenomenon numerically is emphasized. Possible mathematical models for fitting experimental data are classified as statistical, kinetic or empirical and examples of each of these types are given. A possible protocol for analysing cell spreading kinetics and determining goodness of fit and parameter redundancy is presented.

A cell spreading factor, abundant in human placenta, contains fibronectin and fibrinogen.

A salt extract from human normal-term placental extracellular matrix contains material that rapidly adsorbs to glass or tissue-culture plastic and promotes the spreading of fibroblastic and epithelioid cell lines. The cellular response to placental spreading factor (PSF) is characterized and compared with the response to fibronectin. The active component is shown to contain antigenic determinants that cross-react with antibodies to fibronectin and fibrinogen.

Cell surface molecules involved in fibronectin-mediated adhesion. A study using specific antisera.

Attachment and spreading of baby hamster kidney (BHK) fibroblasts to fibronectin-coated surfaces can be inhibited by antibodies, and the (Fab) 2 fragments, prepared against the cells. The antibodies reacted specifically with ricin-binding glycoproteins of the cell surface, the major components having molecular weights of 130-140 K. The antibodies reacted also with mouse fibroblasts (L-cells) and Chinese hamster ovary (CHO)-cells and inhibited their fibronectin-mediated adhesion to inert surfaces.

Cell adhesion on model substrata: threshold effects and receptor modulation.

Trypsinized BHK cells become attached to glass that has been derivatized with a variety of lectins with well-defined specificity for cell-surface carbohydrates. Provided a threshold concentration of glass-immobilized protein is present the cells undergo a transformation to a well-spread morphology. The matrix density of lectins (ricin and concanavalin A) required to trigger this morphological transformation is higher by 10 to 40-fold thant the value determined earlier (Hughes, Pena, Clark & Dourmashkin, 1979) for fibronectin.

Modification of fibroblast surface amines alters receptor-mediated cell spreading on protein-coated substrata but not adsorptive endocytosis.

Fluorescein isothiocyanate (FITC) and other anionic reagents specific for amine groups have previously been shown to inhibit the adhesion and spreading of cultured fibroblasts to fibronectin-coated surfaces (Butters, Devalia, Aplin & Hughes, 1980). Here it is demonstrated that a population of FITC-labelled cells can be separated using flow cytometry into fractions displaying greater and lesser adhesivity at lower and higher fluorescence, respectively, demonstrating that the inhibition is dose-dependent. Glass coverslips covalently derivatized with the lectins ricin and concanavalin A are used to show that the inhibition also occurs in lectinmediated cell adhesion as well as in adhesion to collagen coated with fibronectin and plastic coated with serum or antibody, suggesting that all of these responses share a common, FITC-sensitive component.

Preparation, properties, and applications of carbohydrate conjugates of proteins and lipids.

As a result of the growing awareness of the involvement of the oligosaccharide moieties of glycoproteins and glycolipids in cell surface recognition and binding phenomena, a wide variety of methods have been developed, many quite recently, for preparing glycoconjugates. The chemical methods used for the attachment of sugars and certain hydrophilic polymers (e.g.

Sepharose 4B as a matrix for affinity chromatography. A spin-labelling investigation using nitroxides as model ligands.

Nitroxide spin labels were attached to CNBr-activated Sepharose 4B directly and through oligoglycines and oo-amino-carboxylic acids of varying length. The homogeneity of the carbohydrate environments of directly attached labels was investigated by measuring dipolar interactions between nitroxides as a function of solvation and of spin dilution with a diamagnetic analogue, as well as by electron exchange between the nitroxides and paramagnetic metal ions in solution. Only the exchange experiment revealed any inhomogeneity, suggesting that a small proportion of sites may be less accessible than the majority.

Polypeptide heterogeneity of hamster and calf fibronectins.

The adhesive glycoprotein fibronectin has been isolated from fresh hamster plasma by affinity chromatography on gelatin coupled to Sepharose beads by the method of Engvall & Ruoslahti [Int. J. Cancer (1979) 20, 1-5].

Inhibition of fibronectin-mediated adhesion of hamster fibroblasts to substratum: effects of tunicamycin and some cell surface modifying reagents.

Using baby hamster kidney (BHK) fibroblasts we have studied the effect of tunicamycin, a specific inhibitor of protein glycosylation, on the ability of trypsinized cells to attach and spread onto fibronectin. Tunicamycin inhibited mannose incorporation into total acid-precipitable glycoproteins by at least 95% while glucosamine and leucine incorporation were less or hardly inhibited. Hydrolysis and analysis of [3H]glucosamine-labelled glycoproteins showed that radioactivity incorporated into cells exposed to tunicamycin was present predominantly as galactosamine, presumably present in O-glycosidically linked glycan chains whose assembly is insensitive to the drug.

Spin-labelling of sialic acid in soluble and cell-surface glycoproteins.

A new, mild method is described for spin-labelling sialic acid residues in situ. The procedure involves the formation of C-1 sialamides and has been applied to a serum glycoprotein, a mucin, tissue sections from human colon, and erythrocyte membrane components. The selectivity of the method and its possible applicability to other types of labelling are discussed.