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At least 19 recordsLinked to original sources

Contribution of mucosal chloride to chloride in toad bladder epithelial cells.

Epithelial cells were scraped from the bladders of toads of the species Bufo marinus obtained from the Dominican Republic. These epithelial cells exchanged their chloride virtually completely with 36Cl i; the medium within 60 min. Of this chloride, about 93% came from the serosal medium. The approximately 20 mmole/kg dry wt of chloride which equilibrates with 36Cl in the mucosal medium was still present when choline replaced sodium in the medium in the presence of amiloride (10(-4) M) and was almost all readily removed by rapid washing of the mucosal surface immediately prior to analysis. These observations suggest that little chloride of mucosal origin is truly intracellular. This conclusion is supported by the fact that after vasopressin the increased cellular chloride was not of mucosal origin.

Animals

Cytotoxicity of ricinoleic acid (castor oil) and other intestinal secretagogues on isolated intestinal epithelial cells.

Epithelial cells were isolated from hamster small intestine by a technique of vibration and used to measure cytotoxicity in vitro of certain substances known to stimulate intestinal fluid secretion. These secretagogues have laxative properties and produce mucosal damage in vivo. Compounds tested were ricinoleic acid (caster oil), dioctyl sodium sulfosuccinate, oleic acid, sodium deoxycholate and sodium cholate. Cytotoxicity was assessed by: 1) exclusion of trypan blue; 2) release of intracellular (prelabeled) 51Cr; and 3) inhibition of cellular uptake of 3-O-methylglucose. Ricinoleate produced a dose-dependent (0.1-2.0 mM) cytotoxicity as assessed by all three methods. Oleic acid, a nonhydroxylated analog of ricinoleate, was less potent. The dihydroxy bile acid, deoxycholate, was equipotent with ricinoleate, was less potent. The dihydroxy bile acid, deoxycholate, was equipotent with ricinoleate but its trihydroxy congener, cholate, was less potent. Dioctyl sodium sulfosuccinate had cytotoxicity similar in magnitude to that of ricinoleate and deoxycholate. Cytotoxicity of these agents to isolated cells may relate to their secretory potential in vivo, their abilities to produce structural change at the mucosal surface and their laxative properties.

Animals

Transthoracic aspiration biopsy. The occurrence and significance of alveolar epithelial cells.

Alveolar epithelial cells in pulmonary aspirates sometimes appear with a pronounced nuclear atypism which may give rise to diagnostic difficulties. An analysis has been performed of the occurrence and morphology of alveolar epithelial cells in a series of histologically-verified transthoracic aspiration biopsies not containing clear-cut malignant tumour cells. Alveolar epithelial cells, as well as atypism of these cells, are more often seen in biopsies from histologically malignant cases than in benigh cases. Their presence in a pulmonary aspirate may be due to a chronic obstruction atelectasis. In cases with tumour-like configuration of the alveolar epithelial cells, an open pulmonary biopsy is recommended, as it seems impossible to decide whether such findings represent a neoplastic or a reactive process.

Adolescent

Antiserum to the cell surface of oral epithelial cells: relationship to pemphigus antiserum.

Antisera to human oral epithelial cells were produced in rabbits and found to be specific for epithelial type cells of man, guinea-pig and rabbit. Binding of the antisera to human oral epithelial cells was not affected by pre-incubation in concanavalin A, trypsin or neuraminidase, nor by pre-incubation of the antisera with fetuin. The antisera would appear to differ from pemphigus antiserum.

Animals

Epithelial cell kinetics in the descending colon of the rat. I. The effect of ischaemia-induced epithelial cell loss.

Epithelial cell loss was induced in the descending colon of the rat by temporary ischaemia to investigate whether this would lead to an increase in crypt cell proliferation. Shortly after the temporary ischaemia the number of cells per crypt was markedly reduced, and it was shown that the cell loss occurred mainly from the non-proliferating upper half of the crypt. The number of cells per crypt reached control values again after 24-48 h. There was a marked increase in proliferative activity, as reflected by the labelling index after 3HTdR and by the mitotic index, with peak values at 16 and 24 h after ischaemia. After 48 h the proliferative indices were normal again. The increase in crypt cell proliferation was characterized by an increase in the labelling index as well as in the mitotic index per crypt cell position. No enlargement of the proliferative cell compartment in the crypt was observed. It is most likely then that the increase in crypt cell proliferation was brought about by a shortening of the cell cycle, since the growth fraction in the lower half of the crypt approaches 1.0. The possible implications of the present data for the control of colonic cell proliferation and colonic carcinogenesis are discussed.

Animals

The effects of various mammalian sera on attachment efficiency and thymidine incorporation in primary cultures of mouse mammary epithelial cells.

Mammary epithelial cells from 16- to 17-day pregnant BALB/c mice were cultured in various mammalian sera to determine the kind of serum which stimulates optimal attachment efficiency and thymidine incorporation. Of those sera tested, horse, bovine, lamb, goat and fetal bovine provided the highest attachment efficiency, whereas rat, mouse and human gave the lowest. Rabbit serum stimulated the highest thymidine incorporation into TCA-insoluble material with goat and rat providing the lowest. These results suggest that sera which provide the highest attachment efficiency for primary cultures are not the best stimulants of DNA synthesis and show that an inverse relationship exists between cell attachment and thymidine incorporation for any given type of mammalian serum.

Animals

Adherence of bacterial to vaginal epithelial cells.

Vaginal epithelial cells from healthy women were washed and incubated in tissue culture medium with freshly isolated bacteria of the indigenous vaginal flora and with bacteria of species that have been discussed in conjunction with genital infections. After incubation and washing, the number of bacteria that adhered per cell was determined. The influence on the attachment rate of such factors as variations in the washing procedure, bacterial density, and incubation time was assessed. Lactobacillus acidophilus and other bacterial species that occur in the lower genital tract of healthy women, e.g., some strictly anaerobic species, adhered by significantly lower numbers per cell than Neisseria gonorrhoeae, group B streptococci, and Corynebacterium vaginale. Significantly more freshly isolated gonococci adhered per cell than gonococci that had been passaged on artificial medium. The adherence of gonococci increased with increasing acidity of the test medium.

Adolescent

Relationships between serosal medium potassium concentration and sodium transport in toad urinary bladder. II. Effects of different medium potassium concentrations on epithelial cell composition.

Epithelial cells from hemibladders incubated in potassium-free sodium Ringer's serosal medium lost potassium, both in exchange for serosal sodium and with chloride and water. Cellular sodium of mucosal origin did not change. The loss of cellular potassium, chloride and water closely followed the fall in short-circuit current (SCC). One third as much potassium, chloride and water were lost in 1 mM potassium serosal medium; SCC fell 1/3 as much. Potassium-free choline Ringer's serosal medium abolished the initial increase in SCC and reduced the fall in cellular potassiu, chloride and water and in SCC. Ouabain (10(-2)M) in potassium-free medium prevented the initial increase in SCC and the loss of cellular chloride and water. Ouabain (5 X 10(-4)M) caused loss of cellular potassium in exchange for mucosal and serosal sodium, effects different from those of absence of serosal potassium although SCC was similarly inhibited. Sodium-free mucosal medium abolished SCC and prevented the initial transient of SCC and diminished loss of cellular potassium, chloride and water on removing serosal potassium. When serosal potassium concentration was increased considerably, cells gained potassium, chloride and water, and in 116 mM potassium media, lost sodium of serosal origin. A hypothesis is advanced to explain the transients in SCC on changing serosal potassium concentration. The fall in cellular potassium, not water, probably inhibits sodium transport in media of less than 2 mM potassium.

Animals

Improved protocol for isolation and characterization of human thymic epithelial cells.

Thymic epithelial cells (TECs) play a vital role in the generation of immunocompetent and immunotolerant T cells. The understanding of TEC biology in model animals such as mice has advanced thanks to the cellular and molecular analyses of purified TEC populations, including cortical TECs (cTECs) and medullary TECs (mTECs). On the other hand, studies of human TECs have fallen behind because of the lack of a well-established method for purifying TEC populations. Here, we show that using 0.5 U/mL liberase to digest thymus tissue into single cells is quicker than using collagenase D or 0.1 U/mL liberase. Thymus digestion with 0.5 U/ml liberase preserves CD205 expressed by cTECs. RNA sequencing analysis of TEC populations isolated from 0.5 U/mL liberase-digested human thymic tissue revealed the expression of molecules known as cTEC- and mTEC-specific genes in mouse thymus in each human TEC population. Our study is expected to fuel further cellular and molecular studies of human TECs, including those related to the collapse of immune tolerance in humans.

Flow cytometry

A putative role for cell-cell epithelial contacts in lactose secretion.

Rates of synthesis and secretory release and the maximal requisite intracellular transit time (Tsec) for lactose were measured in vitro for three preparations of lactating guinea pig mammary tissue: tissue slices, mammary epithelial cell (MEC) acini, and mono-dispersed MEC. The Tsec values for tissue slice and acini preparations were similar, lactose required approximately 16 min to pass from its site of synthesis (Golgi) to the extracellular medium. Dispersal of mammary tissue into single cells by collagenase disruption of all cell-cell junctional complexes increased the Tsec value to approximately 25 min but did not alter kinetics of lactose synthesis and secretory release. These data suggest a possible involvement of cell-cell junctional contacts in intracellular transport of lactose.

Animals

Action of vasopressin, ouabain, and cyanide on the volume of isolated toad bladder epithelial cells.

Toad bladder epithelial cells were isolated under mild conditions in a calcium-free medium; they were found to exclude trypan blue, to consume oxygen, and to respond to vasopressin with an increased rate of oxygen consumption. Since isolated toad bladder epithelial cells are mostly spherical in shape, the cell diameter can be accurately measured with an ocular micrometer of an inverted microscope. Epithelial cells swelled by 29+/-3% in the presence of KCN. This cyanide-induced swelling of cells was prevented by amiloride or, alternatively, by replacing NaCl by equiosmotic amounts of mannitol in the Ringer's fluid. Cells incubated in the presence of vasopressin swelled by 10+/-2%. Vasopressin and KCN acted synergistically in enhancing cell volume. Ouabain caused cells to swell by 9+/-2%, and this effect was not additive to the swelling seen with vasopressin. These observations are in accord with the theory of Leaf and his associates, that the predominant effect of vasopressin is to enhance sodium entry into the transporting epithelial cells of the toad urinary bladder.

Amiloride

Cell surface components of intestinal epithelial cells and their relationship to cellular differentiation.

The surfaces of the epithelial cells of the rat small intestine were studied by fluorescence microscopy using various fluorescein isothiocyanate-conjugated lectins. Throughout most of the adult intestine changes were detected in the luminal surfaces as the cells differentiate and move from the crypts to the villi. These changes in cell surface material depend upon the region of intestine in which the villi are located. By the end of the second week of postnatal development the cell surface components detected by the lectins used in this study have appeared at all levels of the small intestine. The maturational and regional differences begin to appear during the third week of postnatal development, and the adult pattern of distribution of surface components is established by 30 days after birth. By eight weeks after proximal jejunal-ileal resectioning or transposition of ileal segments to the jejunum, the epithelial cell surfaces of the relocated ileum had acquired the surface properties characteristic of the jejunal cells. However, jejunal segments transposed to the ileum maintained their original cell surface characteristics and influenced the properties of ileal cell surfaces distally adjacent to the transposed segment. The cell surface material detected by the Ricinus communis (RCAI) lectin was extracted from crypt cells of adult rat jejunum and isolated by affinity chromatography. The isolated material was found to contain at least two components, and its distribution in the jejunum was examined by fluorescence microscopy using antibodies prepared against the isolated material.

Animals

Morphogenesis of porcine rotavirus in porcine kidney cell cultures and intestinal epithelial cells.

The morphogenesis of porcine rotavirus was similar in vitro in porcine kidney (PK) cell cultures and in vivo in porcine epithelial cells as examined by electron microscopy. Infected cells contained cytoplasmic, non-membrane-bound viroplasm and accumulations of virus particles within cisternae of the rough endoplasmic reticulum (RER). Three types of virus particles were noted: double-shelled or complete particles which averaged 77 nm in diam.; single-shelled or naked particles which ranged from 50 to 55 nm in diam.; and electron-dense nucleoids, or cores, 31 to 38 nm in diam. Virus particles acquired outer shells by budding through either matrices of granular, electron-dense viroplasm or membranes of distended RER. Accumulation of numerous single-shelled particles was observed only in PK cell cultures containing a high percentage of infected cells. In these cells, virus release occurred through disruption of the plasma membrane. Tubules, similar in diameter to the single-shelled particles, were observed in the nuclei of a few infected PK cells.

Animals

Telolysomes in cultured iris epithelial cells and in the TVI cell-line.

Iris epithelial cells of adult newts, which are fully differntiated melanocytes and non-dividing, become dedifferentiated and converted into lens cells when put in culture. A recent study shows that this dedifferentiation is based on an autophagic process which is associated with proliferation and mainly affects melanosomes. The present report shows that in primary culture of iris epithelial cells after the majority of melanosomes have disappeared, myelinoid bodies, which are interpreted to be telolysosomes of autophagic nature, appear in high frequencies. This suggest that in these cells autophagy persists after the loss of melanosomes. A possible connection of this type of autophagy with the differentation of lens fiber which occurs in this culture is discussed. In the TVI cell line which is believed to be derived from the same cell type, but devoid of melanosomes, similar myelinoid bodies are a characteristic cell component, suggesting that the tendency for autophagy is inherited in theis cell line.

Animals

Substrate properties influencing ultrastructural differentiation of mammary epithelial cells in culture.

Epithelial cells dissociated from mammary glands of midpregnant mice and cultured with lactogenic hormones on plastic or collagen gel substrates have been shown to vary in their extent of differentiation, as identified by the presence of secretory organelles and accumulation and secretion of casein. Morphological and biochemical differentiation was obtained on floating collagen gels. At least four unique factors provided by the floating collagen gel substrates are not found on plastic substrates: access of nutrients to basolateral cell surfaces, close proximity of cells to the medium surface and gas phase, interaction of epithelial cells with stromal elements, and substrate flexibility permitting cell shape change. In this study, we have attempted to assess the relative contributions of these factors in the ultrastructural differentiation of mammary cells in culture. None of these factors alone is responsible for the differentiation achieved when all are present. The novel aspect of this research is the identification of the cells' apparent requirements for basolateral access to nutrients and for freedom to assume a preferred shape in order to achieve differentiation.

Animals