Isolation, in vitro cultivation, and electron microscopy of normal and malignant prostatic epithelial cells from the Copenhagen rat

1980 ◽  
Vol 8 (3) ◽  
pp. 139-152 ◽  
Author(s):  
Frederick A. Feuchter ◽  
David R. Rowley ◽  
Paul M. Heidger
Keyword(s):  
Electron Microscopy ◽  
Epithelial Cells ◽  
In Vitro Cultivation ◽  
Copenhagen Rat

scholarly journals Lens metabolic cooperation: a study of mouse lens transport and permeability visualized with freeze-substitution autoradiography and electron microscopy.

1980 ◽  
Vol 86 (2) ◽  
pp. 576-589 ◽  
Author(s):  
D A Goodenough ◽  
J S Dick ◽  
J E Lyons
Keyword(s):  
Electron Microscopy ◽  
Epithelial Cells ◽  
Freeze Fracture ◽  
Freeze Substitution ◽  
Species Variation ◽  
Metabolic Cooperation ◽  
Zonulae Occludentes ◽  
Lens Fibers ◽  
Mouse Lens

Transport of metabolites is demonstrated between compartments of the adult mouse lens by freeze-substitution autoradiography. In vivo patterns of lysine incorporation are compared with in vitro patterns of lysine, glucose, uridine, and deoxyglucose incorporation. Intracellular and extracellular distributions of tritiated metabolites are determined by comparison of transported substrates with the nontransported molecules of similar molecular size: mannitol and sucrose. The permeability of the lens intercellular spaces is probed with Procion Yellow at the level of fluorescence microscopy, and with horseradish peroxidase at the electron microscope level. Freeze-fracture electron microscopy reveals gap junctions between epithelial cells, between lens fibers, and between epithelial cells and lens fibers. Zonulae occludentes (tight junctions) are not routinely observed between epithelial cells in the mouse. This latter result is subject to species variation, however, since zonulae occludentes are abundant between chicken epithelial cells. The permeability results suggest that the lens cells are capable of metabolic cooperation, mediated by an extensive gap junction network.

scholarly journals Genes encoding proteins regulating fatty acid metabolism and cellular response to lipids are differentially expressed in porcine luminal epithelium during long-term culture

2019 ◽  
Vol 7 (2) ◽  
pp. 58-65
Author(s):  
Magdalena Kulus ◽  
Blanka Borowiec ◽  
Małgorzata Popis ◽  
Piotr Celichowski ◽  
Michal Jeseta ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Epithelial Cells ◽  
Cellular Response ◽  
Molecular Mechanisms ◽  
In Vitro Cultivation ◽  
Beta Oxidation ◽  
Physiological Processes ◽  
Genes Encoding ◽  
Fatty Acid Beta Oxidation

AbstractAmong many factors, the epithelium lining the oviductal lumenis very important for the development of the oocyte and its subsequent fertilization. The oviductal epithelium is characterized by the presence of ciliary cells, supporting the movement of cumulus-oocyte complexes towards the uterus. By interacting with the semen, the epithelium of the fallopian tube makes the sperm acquire the ability to fertilize. So far, the exact molecular mechanisms of these changes have not been known. Hence, understanding the metabolism of oviduct epithelial cells and the level of expression of individual groups of genes seems to be a way to deepen the knowledge about the broadly understood reproduction.In our research, we decided to culture oviductal epithelial cells (OECs) in vitro for a long period of time. After 24h, 7, 15 and 30 days, the OECs were harvested, with their RNA isolated. Transcriptomic changes were analyzed using microarrays. The “cellular response to lipid” group was represented by the following genes: MUC1, CYP24A1, KLF4, IL24, SNAI2, CXCL10, PPARD, TNC, ABCA10, while the genes belonging to the “cellular lipid metabolic processes” were: LIPG, ARSK, ACADL, FADS3, P2RX7, ACSS2, PPARD, KITLG, SPTLC3, ERBB3, KLF4, CRABP2. Additionally, PPARD and ACADL were members of the “fatty acid beta-oxidation” ontology group. Our study describes genes that are not directly related to fertility processes. However, significant changes in their expression in in vitro cultured OECs may indicate their usefulness as markers of OECs’ physiological processes.Running title: Fatty acids changes in porcine oviductal epithelial cells in in vitro cultivation

scholarly journals HOST-PARASITE INTERACTION IN THE RAT RENAL PELVIS

1974 ◽  
Vol 140 (6) ◽  
pp. 1696-1711 ◽  
Author(s):  
Fredric J. Silverblatt
Keyword(s):  
Electron Microscopy ◽  
Epithelial Cells ◽  
Female Rats ◽  
Mucosal Barrier ◽  
Type Iii ◽  
Type Iv ◽  
Predominant Type ◽  
Initial Interaction ◽  
Host Parasite

The initial interaction between bacteria and the renal pelvic epithelium may determine whether intrarenal infection occurs. A model of retrograde pyelonephritis was employed to study these events by electron microscopy. Female rats received an intravesicular inoculation of a 0.5-ml suspension of Proteus mirabilis containing 108 organisms. At intervals after inoculation, the kidneys were fixed by intravascular perfusion and the tissues were prepared for electron microscopy. During the first 24 h, increasing numbers of bacteria were seen to be attached by pili to the renal pelvic epithelial cells. The organism appeared to cross the mucosal barrier by several mechanisms: (a) penetration into the cytoplasm of intact epithelial cells, (b) passage between epithelial cells that were separated by excessive hydrostatic pressure generated during bladder inoculation, (c) passage across necrotic regions of the pelvis, and (d) translocation to the cortex by calicotubular backflow. Whereas at inoculation bacteria possessed pili 40 Å in diameter (type III pili) 24 h after reflux, the predominant type of pili measured 70 A in thickness (type IV pili). Repetitive subculture induced a similar transition in vitro. To assess the influence of pili type on virulence in this model, 80 rats were challenged with either type III or type IV pilated organisms and the frequency of rats with cortical abscesses were compared at 1 wk. A significantly greater number of rats inoculated with type IV pilated Proteus manifested macroscopic evidence of infection. These results suggest that pili play a role in the pathogenesis of ascending pyelonephritis.

scholarly journals Cell Wall-associated Protein A as a Tool for Immunolocalization of Staphylococcus aureus in Infected Human Airway Epithelium

2000 ◽  
Vol 48 (4) ◽  
pp. 523-533 ◽  
Author(s):  
Emmanuel Mongodin ◽  
Odile Bajolet ◽  
Jocelyne Hinnrasky ◽  
Edith Puchelle ◽  
Sophie de Bentzmann
Keyword(s):  
Electron Microscopy ◽  
Staphylococcus Aureus ◽  
Epithelial Cells ◽  
Airway Epithelium ◽  
Protein A ◽  
Staphylococcal Protein A ◽  
Basal Cells ◽  
Bronchial Diseases

Staphylococcus aureus is a common human pathogen involved in non-bronchial diseases and in genetic and acquired bronchial diseases. In this study, we applied an immunolabeling approach for in vivo and in vitro detection of S. aureus, based on the affinity of staphylococcal protein A (SpA) for the Fc region of immunoglobulins, especially IgG. Most strains of S. aureus, including clinical strains, can be detected with this labeling technique. The approach can be used for detection and localization with transmission electron microscopy or light-fluorescence microscopy of S. aureus in infected tissues such as human bronchial tissue from cystic fibrosis (CF) patients. The methodology can also be applied to cell culture models with the aim of characterizing bacterial adherence to epithelial cells in backscattered electron imaging with scanning electron microscopy. Application to the study of S. aureus adherence to airway epithelium showed that the bacteria did not adhere in vivo to intact airway epithelium. In contrast, bacteria adhered to the basolateral plasma membrane of columnar cells, to basal cells, to the basement membrane and were identified beneath the lamina propria when the epithelium was injured and remodeled, or in vitro when the epithelial cells were dedifferentiated.

scholarly journals In Vitro Cultivation and Electron Microscopy Characterization of Trachipleistophora anthropophthera Isolated From the Cornea of an AIDS Patient

2005 ◽  
Vol 52 (3) ◽  
pp. 179-190 ◽  
Author(s):  
SANDRA I. JUAREZ ◽  
CHATURONG PUTAPORNTIP ◽  
SOMCHAI JONGWUTIWES ◽  
AKITOYO ICHINOSE ◽  
TETSUO YANAGI ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
In Vitro Cultivation ◽  
Microscopy Characterization

scholarly journals Molting, Ecdysis, and Reproduction of Trichinella spiralis Are Supported In Vitro by Intestinal Epithelial Cells

2002 ◽  
Vol 70 (4) ◽  
pp. 1853-1859 ◽  
Author(s):  
L. F. Gagliardo ◽  
C. S. McVay ◽  
J. A. Appleton
Keyword(s):  
Life Cycle ◽  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
Trichinella Spiralis ◽  
Mature Male ◽  
In Vitro Cultivation ◽  
Low Density ◽  
Intestinal Epithelial ◽  
Prolonged Culture

ABSTRACT Trichinella spiralis is an obligate parasite of animals that has an unusual intracellular life cycle. Investigation of parasitism at the cellular and molecular levels has been challenging because of a shortage of tools for in vitro cultivation of T. spiralis. We have found that T. spiralis larvae molt, ecdyse, develop to adulthood, and reproduce when they are inoculated onto cultured intestinal epithelial cells. Initially, larvae invade and migrate through cells in a monolayer (T. ManWarren, L. Gagliardo, J. Geyer, C. McVay, S. Pearce-Kelling, and J. Appleton, Infect. Immun. 65:4806-4812, 1997). During prolonged culture in Caco-2 epithelial cells, L1 larvae molted and ecdysed with efficiencies as high as 50%. Molting and ecdysis in vitro required entry of the parasite into cells; conditions that prevented entry into cells also prevented ecdysis. When larvae were inoculated at a low density and cultured for 5 to 9 days, as many as 50% of the larvae developed to adult stages. Low numbers of mature male worms with copulatory appendages were observed in these cultures. The majority of worms that survived for five or more days were unfertilized females. Low-density cultures supported development of female worms with embryos at rates of 4 to 5%. These results show that the intestinal life cycle of T. spiralis can be supported entirely by host epithelial cells. Our model should allow more detailed investigation of intracellular parasitism by T. spiralis.

scholarly journals A Double, Long Polar Fimbria Mutant of Escherichia coli O157:H7 Expresses Curli and Exhibits ReducedIn VivoColonization

2012 ◽  
Vol 80 (3) ◽  
pp. 914-920 ◽  
Author(s):  
Sonja J. Lloyd ◽  
Jennifer M. Ritchie ◽  
Maricarmen Rojas-Lopez ◽  
Carla A. Blumentritt ◽  
Vsevolod L. Popov ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Escherichia Coli ◽  
Epithelial Cells ◽  
Rabbit Model ◽  
Intestinal Colonization ◽  
Content Type ◽  
Regulatory Pathways ◽  
E Coli ◽  
Reverse Transcription Pcr

Escherichia coliO157:H7 causes food and waterborne enteric infections that can result in hemorrhagic colitis and life-threatening hemolytic uremic syndrome. Intimate adherence of the bacteria to intestinal epithelial cells is mediated by intimin, butE. coliO157:H7 also possess several other putative adhesins, including curli and two operons that encode long polar fimbriae (Lpf). To assess the importance of Lpf for intestinal colonization, we performed competition experiments betweenE. coliO157:H7 and an isogenic ΔlpfA1ΔlpfA2double mutant in the infant rabbit model. The mutant was outcompeted in the ileum, cecum, and midcolon, suggesting that Lpf contributes to intestinal colonization. In contrast, the ΔlpfA1ΔlpfA2mutant showed increased adherence to colonic epithelial cellsin vitro. Transmission electron microscopy revealed curli-like structures on the surface of the ΔlpfA1ΔlpfA2mutant, and the presence of curli was confirmed by Congo red binding, immunogold-labeling electron microscopy, immunoblotting, and quantitative real-time reverse transcription-PCR (qRT-PCR) measuringcsgAexpression. However, deletion ofcsgA, which encodes the major curli subunit, does not appear to affect intestinal colonization. In addition to suggesting that Lpf can contribute to EHEC intestinal colonization, our observations indicate that the regulatory pathways governing the expression of Lpf and curli are interdependent.

scholarly journals The genes regulating maintenance of cellular protein location are differentially expressed in porcine epithelial oviductal cells during longterm in vitro cultivation

2019 ◽  
Vol 7 (2) ◽  
pp. 77-85 ◽  
Author(s):  
Katarzyna Stefańska ◽  
Ievgenia Kocherova ◽  
Sandra Knap ◽  
Magdalena Kulus ◽  
Piotr Celichowski ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Differentially Expressed Genes ◽  
Cellular Protein ◽  
Early Embryo ◽  
Female Reproductive Tract ◽  
Differentially Expressed ◽  
In Vitro Cultivation ◽  
Protein Location

AbstractThe oviduct is a part of female reproductive tract that is essential for successful fertilization and early embryo development. It is lined with epithelium consisting of two types of cells: ciliated and secretory. The primary function of ciliated oviductal epithelial cells (OECs) is to support the transport of gametes and embryos through the ovary, whereas secretory OECs produce components of the oviductal fluid. Undoubtedly, the oviductal epithelium plays a major part in the early aspects of pregnancy development, from providing an optimal environment for gametes and embryos to supporting fertilization. Therefore, our aim was to gain a better insight into the genetic changes underlying function of these cells. We have harvested OECs from crossbred gilts (n=45), at the age of about nine months and which displayed two regular estrous cycles, and established long-term primary culture of porcine OECs. Microarray analysis was utilized to determine differentially expressed genes during day 1, 7, 15 and 30 of cultivation, with our results revealing54 differentially expressed genes belonging to three ontology groups: „maintenance of location”, „maintenance of protein location” and „maintenance of protein location in cell”. Since the biochemistry and morphology of epithelial cells may change during long term cultivation, we conclude that our results are a reflection of these changes and help to shed a light on porcine OECs properties in in vitro environment.Running title: Maintenance of cellular protein location in porcine epithelial oviductal cells

UPTAKE OF 131I BY THE THYROID OF THE INFANTILE RAT AFTER PROLONGED IN VITRO CULTIVATION

1955 ◽  
Vol 12 (4) ◽  
pp. 227-NP ◽  
Author(s):  
MIROSLAVA R. PAVLOVIĆ
Keyword(s):  
Epithelial Cells ◽  
Radioactive Iodine ◽  
In Vitro Cultivation ◽  
Thyroid Glands

SUMMARY The thyroid glands of infantile rats cultivated in vitro for up to 2 months retain their capacity to concentrate radioactive iodine selectively. It is established autoradiographically that the 131I taken up by the cultivated gland is localized both in the epithelial cells and colloid of healthy thyroid follicles.

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